Thiophosphate nucleosides for the treatment of HCV

ABSTRACT

Provided herein are compounds, compositions and methods for the treatment of Flaviviridae infections, including HCV infections. In certain embodiments, the compounds are according to Formula 2001: 
                         
where PD, Base, R A  and R B  are as provided herein. In certain embodiments, compounds and compositions of nucleoside derivatives are disclosed, which can be administered either alone or in combination with other anti-viral agents.

FIELD

Provided herein are compounds, methods, and pharmaceutical compositionsfor use in treatment of viral infections, including hepatitis C virusinfections in hosts in need thereof. In certain embodiments,thiophosphate nucleosides are provided which display remarkable efficacyand bioavailability for the treatment of, for example, HCV infection ina human.

BACKGROUND

The hepatitis C virus (HCV) is the leading cause of chronic liverdisease worldwide. (Boyer, N. et al., J. Hepatol. 32:98-112, 2000). HCVcauses a slow growing viral infection and is the major cause ofcirrhosis and hepatocellular carcinoma (Di Besceglie, A. M. and Bacon,B. R., Scientific American, October: 80-85, 1999; Boyer, N. et al., J.Hepatol. 32:98-112, 2000). It is estimated there are about 130-170million people with chronic hepatitis C virus infection, and there areabout 350,000 deaths from hepatitis C-related liver diseases each year(Hepatitis C Fact Sheet, World Health Organization Fact Sheet No. 164,June 2011). Cirrhosis caused by chronic hepatitis C infection accountsfor 8,000-12,000 deaths per year in the United States, and HCV infectionis the leading indication for liver transplantation.

HCV infection becomes chronic in about 75% of cases, with many patientsinitially being asymptomatic. The first symptoms of HCV infection areoften those of chronic liver disease. About 20 to 30% of patients withchronic hepatitis due to HCV develop cirrhosis, although this may takedecades. Development of cirrhosis due to HCV also increases the risk ofhepatocellular cancer (The Merck Manual Online, Chronic Hepatitis,available atwww.merckmanuals.com/professional/hepatic_and_biliary_disorders/hepatitis/chronic_hepatitis.html, last revision February 2007).

In light of the fact that HCV infection has reached epidemic levelsworldwide, and has tragic effects on the infected patient, there remainsa strong need to provide new effective pharmaceutical agents to treathepatitis C that have low toxicity to the host. Further, given therising threat of other flaviviridae infections, there remains a strongneed to provide new effective pharmaceutical agents that have lowtoxicity to the host. Therefore, there is a continuing need foreffective treatments of flavivirus infections and HCV infections.

SUMMARY

Provided herein are compounds useful, for example, for the treatment offlavivirus infections such as HCV infections. The compounds arethiophosphate nucleosides. In certain embodiments the thiophosphatenucleosides display remarkable efficacy or bioavailability, or both, forthe treatment of, for example, HCV infection in a human.

In certain embodiments, the compounds provided herein are useful in theprevention and treatment of Flaviviridae infections and other relatedconditions such as anti-Flaviviridae antibody positive andFlaviviridae-positive conditions, chronic liver inflammation caused byHCV, cirrhosis, fibrosis, acute hepatitis, fulminant hepatitis, chronicpersistent hepatitis, and fatigue. These compounds or formulations canalso be used prophylactically to prevent or retard the progression ofclinical illness in individuals who are anti-Flaviviridae antibody orFlaviviridae-antigen positive or who have been exposed to aFlaviviridae. In particular embodiments, the Flaviviridae is hepatitisC. In certain embodiments, the compounds are used to treat any virusthat replicates through an RNA-dependent RNA polymerase.

A method for the treatment of a Flaviviridae infection in a host,including a human, is also provided that includes administering aneffective amount of a compound provided herein, administered eitheralone or in combination or alternation with another anti-Flaviviridaeagent, optionally in a pharmaceutically acceptable carrier.

In certain embodiments, provided herein are compounds according toFormula 2001:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein:

-   -   Base is a nucleobase;    -   R^(A) is hydroxyl or halo;    -   R^(B) is methyl or halo;    -   PD is

-   -   X is alkyl, alkoxycarbonylalkyl,        (alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,        heterocycloalkyl, hydantoinylalkyl, or aryl;    -   Y is —NR¹R² or an N-linked or O-linked amino acid residue, or a        derivative thereof;    -   Z is alkyl, alkoxycarbonyl, or alkoxyalkylcarbonyl;    -   A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;    -   R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl,        or heteroarylalkyl;    -   R² is hydrogen or alkyl;    -   R is hydrogen or alkyl; and    -   each of R¹⁰, R¹¹, and R¹² is independently hydrogen or alkyl; or        R¹⁰ and R¹¹, together with the carbon atoms to which they are        attached, combine to form a five-, six-, or seven-membered ring.

In certain embodiments, provided herein are compounds according toFormula (I):

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein: Base is anucleobase; R^(A) is hydroxyl or halo; R^(B) is methyl or halo; PD is

X is alkyl, alkoxycarbonylalkyl,(alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,heterocycloalkyl, hydantoinylalkyl, or aryl; Y is —NR¹R² or an N-linkedamino acid, or ester thereof; Z is alkyl, alkoxycarbonyl oralkoxyalkylcarbonyl; A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl, orheteroarylalkyl; R² is hydrogen or alkyl; R is hydrogen or alkyl; andeach of R¹⁰, R¹¹, and R¹² is independently hydrogen or alkyl; or R¹⁰ andR¹¹, together with the carbon atoms to which they are attached, combineto form a five-, six-, or seven-membered ring.

In one aspect, the compounds provided herein are provided oradministered in combination with a second therapeutic agent, such as oneuseful for the treatment or prevention of HCV infections. Exemplarysecond therapeutic agents are provided in detail elsewhere herein.

In another aspect, provided herein are pharmaceutical compositions,single unit dosage forms, and kits suitable for use in treating orpreventing disorders such as HCV infections which comprise atherapeutically or prophylactically effective amount of a compoundprovided herein, e.g., of Formula 2001, I-XXIX, 1-23byii and 1001-1002and a therapeutically or prophylactically effective amount of a secondtherapeutic agent such as one useful for the treatment or prevention ofHCV infections.

In certain embodiments, a method of treatment of a liver disorder isprovided comprising administering to an individual in need thereof atreatment effective amount of a thiophosphate nucleoside compound.

Flaviviridae which can be treated are, e.g., discussed generally inFields Virology, Fifth Ed., Editors: Knipe, D. M., and Howley, P. M.,Lippincott Williams & Wilkins Publishers, Philadelphia, Pa., Chapters33-35, 2006. In a particular embodiment of the invention, theFlaviviridae is HCV. In an alternate embodiment, the Flaviviridae is aflavivirus or pestivirus. In certain embodiments, the Flaviviridae canbe from any class of Flaviviridae. In certain embodiments, theFlaviviridae is a mammalian tick-borne virus. In certain embodiments,the Flaviviridae is a seabird tick-borne virus. In certain embodiments,the Flaviviridae is a mosquito-borne virus. In certain embodiments, theFlaviviridae is an Aroa virus. In certain embodiments, the Flaviviridaeis a Dengue virus. In certain embodiments, the Flaviviridae is aJapanese encephalitis virus. In certain embodiments, the Flaviviridae isa Kokobera virus. In certain embodiments, the Flaviviridae is a Ntayavirus. In certain embodiments, the Flaviviridae is a Spondweni virus. Incertain embodiments, the Flaviviridae is a Yellow fever virus. Incertain embodiments, the Flaviviridae is a Entebbe virus. In certainembodiments, the Flaviviridae is a Modoc virus. In certain embodiments,the Flaviviridae is a Rio Bravo virus.

Specific flaviviruses which can be treated include, without limitation:Absettarov, Aedes, Alfuy, Alkhurma, Apoi, Aroa, Bagaza, Banzi, Bukalasabat, Bouboui, Bussuquara, Cacipacore, Calbertado, Carey Island, Cellfusing agent, Cowbone Ridge, Culex, Dakar bat, Dengue 1, Dengue 2,Dengue 3, Dengue 4, Edge Hill, Entebbe bat, Gadgets Gully, Hanzalova,Hypr, Ilheus, Israel turkey meningoencephalitis, Japanese encephalitis,Jugra, Jutiapa, Kadam, Kamiti River, Karshi, Kedougou, Kokobera,Koutango, Kumlinge, Kunjin, Kyasanur Forest disease, Langat, Loupingill, Meaban, Modoc, Montana myotis leukoencephalitis, Murray valleyencephalitis, Nakiwogo, Naranjal, Negishi, Ntaya, Omsk hemorrhagicfever, Phnom-Penh bat, Powassan, Quang Binh, Rio Bravo, Rocio, RoyalFarm, Russian spring-summer encephalitis, Saboya, St. Louisencephalitis, Sal Vieja, San Perlita, Saumarez Reef, Sepik, Sokuluk,Spondweni, Stratford, Tembusu, Tick-borne encephalitis, Turkish sheepencephalitis, Tyuleniy, Uganda S, Usutu, Wesselsbron, West Nile,Yaounde, Yellow fever, Yokose, and Zika.

Pestiviruses which can be treated are discussed generally in FieldsVirology, Fifth Ed., Editors: Knipe, D. M., and Howley, P. M.,Lippincott Williams & Wilkins Publishers, Philadelphia, Pa., Chapters33-35, 2006. Specific pestiviruses which can be treated include, withoutlimitation: bovine viral diarrhea virus (“BVDV”), classical swine fevervirus (“CSFV,” also called hog cholera virus), and border disease virus(“BDV”).

DESCRIPTION OF EXEMPLARY EMBODIMENTS

Provided herein are compounds, compositions and methods useful fortreating liver disorders such as HCV infection in a subject. Furtherprovided are dosage forms useful for such methods.

DEFINITIONS

When referring to the compounds provided herein, the following termshave the following meanings unless indicated otherwise. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as is commonly understood by one of ordinary skill in the art.In the event that there is a plurality of definitions for a term herein,those in this section prevail unless stated otherwise.

The term “alkyl,” as used herein, unless otherwise specified, refers toa saturated straight or branched hydrocarbon. In certain embodiments,the alkyl group is a primary, secondary, or tertiary hydrocarbon. Incertain embodiments, the alkyl group includes one to ten carbon atoms,i.e., C₁ to C₁₀ alkyl. In certain embodiments, the alkyl group ismethyl, CF₃, CCl₃, CFCl₂, CF₂Cl, ethyl, CH₂CF₃, CF₂CF₃, propyl,isopropyl, butyl, isobutyl, secbutyl, t-butyl, pentyl, isopentyl,neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, or2,3-dimethylbutyl. The term includes both substituted and unsubstitutedalkyl groups, including halogenated alkyl groups. In certainembodiments, the alkyl group is a fluorinated alkyl group. Non-limitingexamples of moieties with which the alkyl group can be substitutedinclude, but not limited to, halogen (fluoro, chloro, bromo, or iodo),hydroxyl, carbonyl, sulfanyl, amino, alkylamino, arylamino, alkoxy,aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid,phosphate, or phosphonate, either unprotected, or protected asnecessary, as known to those skilled in the art, for example, as taughtin Greene, et al., Protective Groups in Organic Synthesis, John Wileyand Sons, Second Edition, 1991, hereby incorporated by reference.

The term “lower alkyl,” as used herein, and unless otherwise specified,refers to a saturated straight or branched hydrocarbon having one to sixcarbon atoms, i.e., C₁ to C₆ alkyl. In certain embodiments, the loweralkyl group is a primary, secondary, or tertiary hydrocarbon. The termincludes both substituted and unsubstituted moieties.

The term “upper alkyl,” as used herein, and unless otherwise specified,refers to a saturated straight or branched hydrocarbon having seven tothirty carbon atoms, i.e., C₇ to C₃₀ alkyl. In certain embodiments, theupper alkyl group is a primary, secondary, or tertiary hydrocarbon. Theterm includes both substituted and unsubstituted moieties.

The term “cycloalkyl,” as used herein, unless otherwise specified,refers to a saturated cyclic hydrocarbon. In certain embodiments, thecycloalkyl group may be a saturated, and/or bridged, and/or non-bridged,and/or a fused bicyclic group. In certain embodiments, the cycloalkylgroup includes three to ten carbon atoms, i.e., C₃ to C₁₀ cycloalkyl. Insome embodiments, the cycloalkyl has from 3 to 15 (C₃₋₁₅), from 3 to 10(C₃₋₁₀), or from 3 to 7 (C₃₋₇) carbon atoms. In certain embodiments, thecycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl,bicyclo[2.2.1]heptyl, decalinyl, or adamantyl. The term includes bothsubstituted and unsubstituted cycloalkyl groups, including halogenatedcycloalkyl groups. In certain embodiments, the cycloalkyl group is afluorinated cycloalkyl group. Non-limiting examples of moieties withwhich the cycloalkyl group can be substituted include, but not limitedto, halogen (fluoro, chloro, bromo, or iodo), hydroxyl, carbonyl,sulfanyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate,either unprotected, or protected as necessary.

“Alkylene” refers to divalent saturated aliphatic hydrocarbon groupsparticularly having from one to eleven carbon atoms which can bestraight-chained or branched. In certain embodiments, the alkylene groupcontains 1 to 10 carbon atoms. The term includes both substituted andunsubstituted moieties. This term is exemplified by groups such asmethylene (—CH₂—), ethylene (—CH₂CH₂—), the propylene isomers (e.g.,—CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like. The term includeshalogenated alkylene groups. In certain embodiments, the alkylene groupis a fluorinated alkylene group. Non-limiting examples of moieties withwhich the alkylene group can be substituted include, but not limited to,halogen (fluoro, chloro, bromo, or iodo), hydroxyl, carbonyl, sulfanyl,amino, alkylamino, alkylaryl, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, and phosphonate,either unprotected, or protected as necessary.

“Alkenyl” refers to monovalent olefinically unsaturated hydrocarbongroups, in certain embodiment, having up to about 11 carbon atoms, from2 to 8 carbon atoms, or from 2 to 6 carbon atoms, which can bestraight-chained or branched and having at least 1 or from 1 to 2 sitesof olefinic unsaturation. The term includes both substituted andunsubstituted moieties. Exemplary alkenyl groups include ethenyl (i.e.,vinyl or —CH═CH₂), n-propenyl (—CH₂CH═CH₂), isopropenyl (—C(CH₃)═CH₂),and the like. The term includes halogenated alkenyl groups. In certainembodiments, the alkenyl group is a fluorinated alkenyl group.Non-limiting examples of moieties with which the alkenyl group can besubstituted include, but not limited to, halogen (fluoro, chloro, bromo,or iodo), hydroxyl, carbonyl, sulfanyl, amino, alkylamino, arylamino,alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid,phosphate, or phosphonate, either unprotected, or protected asnecessary.

The term “cycloalkenyl,” as used herein, unless otherwise specified,refers to an unsaturated cyclic hydrocarbon. In certain embodiments,cycloalkenyl refers to mono- or multicyclic ring systems that include atleast one double bond. In certain embodiments, the cycloalkenyl groupmay be a bridged, non-bridged, and/or a fused bicyclic group. In certainembodiments, the cycloalkyl group includes three to ten carbon atoms,i.e., C₃ to C₁₀ cycloalkyl. In some embodiments, the cycloalkenyl hasfrom 3 to 7 (C₃₋₁₀), or from 4 to 7 (C₄₋₇) carbon atoms. The termincludes both substituted and unsubstituted cycloalkenyl groups,including halogenated cycloalkenyl groups. In certain embodiments, thecycloalkenyl group is a fluorinated cycloalkenyl group. Non-limitingexamples of moieties with which the cycloalkenyl group can besubstituted include, but not limited to, halogen (fluoro, chloro, bromo,or iodo), hydroxyl, carbonyl, sulfanyl, amino, alkylamino, arylamino,alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid,phosphate, or phosphonate, either unprotected, or protected asnecessary.

“Alkenylene” refers to divalent olefinically unsaturated hydrocarbongroups, in certain embodiments, having up to about 11 carbon atoms orfrom 2 to 6 carbon atoms which can be straight-chained or branched andhaving at least 1 or from 1 to 2 sites of olefinic unsaturation. Thisterm is exemplified by groups such as ethenylene (—CH═CH—), thepropenylene isomers (e.g., —CH═CHCH₂— and —C(CH₃)═CH— and —CH═C(CH₃)—)and the like. The term includes both substituted and unsubstitutedalkenylene groups, including halogenated alkenylene groups. In certainembodiments, the alkenylene group is a fluorinated alkenylene group.Non-limiting examples of moieties with which the alkenylene group can besubstituted include, but not limited to, halogen (fluoro, chloro, bromo,or iodo), hydroxyl, carbonyl, sulfanyl, amino, alkylamino, arylamino,alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid,phosphate, or phosphonate, either unprotected, or protected asnecessary.

“Alkynyl” refers to acetylenically unsaturated hydrocarbon groups, incertain embodiments, having up to about 11 carbon atoms or from 2 to 6carbon atoms which can be straight-chained or branched and having atleast 1 or from 1 to 2 sites of alkynyl unsaturation. Non-limitingexamples of alkynyl groups include acetylenic, ethynyl (—C≡CH),propargyl (—CH₂C≡CH), and the like. The term includes both substitutedand unsubstituted alkynyl groups, including halogenated alkynyl groups.In certain embodiments, the alkynyl group is a fluorinated alkynylgroup. Non-limiting examples of moieties with which the alkynyl groupcan be substituted include, but not limited to, halogen (fluoro, chloro,bromo, or iodo), hydroxyl, carbonyl, sulfanyl, amino, alkylamino,arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate,phosphonic acid, phosphate, or phosphonate, either unprotected, orprotected as necessary.

The term “aryl,” as used herein, and unless otherwise specified, refersto a substituent derived from an aromatic ring. In an embodiment, anaryl group is a C₆-C₁₂ aryl group. In an embodiment, an aryl group isphenyl, biphenyl, or naphthyl. The term includes both substituted andunsubstituted moieties. An aryl group can be substituted with anydescribed moiety, including, but not limited to, one or more moietiesselected from halogen (fluoro, chloro, bromo, or iodo), alkyl,haloalkyl, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy,nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, orphosphonate, either unprotected, or protected as necessary, as known tothose skilled in the art, for example, as taught in Greene, et al.,Protective Groups in Organic Synthesis, John Wiley and Sons, SecondEdition, 1991.

“Alkoxy” refers to the group —OR′ where R′ is alkyl or cycloalkyl.Alkoxy groups include, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy,1,2-dimethylbutoxy, and the like.

“Alkoxycarbonyl” refers to a radical —C(O)-alkoxy where alkoxy is asdefined herein.

“Alkoxyalkylcarbonyl” refers to a radical —C(O)-alkyl-alkoxy wherealkoxy and alkyl are as defined herein.

“Alkoxycarbonylalkyl” refers to a radical -alkyl-C(O)-alkoxy wherealkoxy and alkyl are as defined herein.

“Alkoxycarbonylamino” refers to a radical -amino-C(O)-alkoxy wherealkoxy and amino are as defined herein.

As used herein, “(alkoxycarbonyl)(alkoxycarbonylamino)alkyl” refers toan alkyl radical substituted with both an alkoxycarbonyl and analkoxycarbonylamino group, where “alkoxycarbonyl” and“alkoxycarbonylamino” are as described herein. In an embodiment, theterm refers to a radical of formula

wherein n is an integer selected over the range of 1-10, A^(A) is—C(O)—O—R¹⁰⁰, A^(N) is —NH—C(O)—O—R¹⁰¹, and each of R¹⁰⁰ and R¹⁰¹ isindependently lower alkyl. In an embodiment, each of R¹⁰⁰ and R¹⁰¹ isindependently C₁-C₅ alkyl.

“Amino” refers to the group —NR^(1′)R^(2′) or —NR^(1′)—, wherein R^(1′)and R^(2′) are independently selected from hydrogen, alkyl andcycloalkyl.

“Amino alcohol” refers to the radical —NHLOH, wherein L is alkylene.

“Carboxyl” or “carboxy” refers to the radical —C(O)OH.

The term “alkylamino” or “arylamino” refers to an amino group that hasone or two alkyl or aryl substituents, respectively. In certainembodiments, the alkyl substituent is upper alkyl. In certainembodiments, the alkyl substituent is lower alkyl. In anotherembodiment, the alkyl, upper alkyl, or lower alkyl is unsubstituted.

“Halogen” or “halo” refers to chloro, bromo, fluoro, or iodo.

“Thioalkoxy” refers to the group —SR′ where R′ is alkyl or cycloalkyl.

The term “heterocyclo” or “heterocyclic” refers to a monovalentmonocyclic non-aromatic ring system and/or multicyclic ring system thatcontains at least one non-aromatic ring, wherein one or more of thenon-aromatic ring atoms are heteroatoms independently selected from O,S, or N; and the remaining ring atoms are carbon atoms. In certainembodiments, the heterocyclo or heterocyclic group has from 3 to 20,from 3 to 15, from 3 to 10, from 3 to 8, from 4 to 7, or from 5 to 6ring atoms. Heterocyclo groups are bonded to the rest of the moleculethrough the non-aromatic ring. In certain embodiments, the heterocyclois a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, whichmay include a fused or bridged ring system, and in which the nitrogen orsulfur atoms may be optionally oxidized, the nitrogen atoms may beoptionally quaternized, and some rings may be partially or fullysaturated, or aromatic. The heterocyclo may be attached to the mainstructure at any heteroatom or carbon atom which results in the creationof a stable compound. Examples of such heterocyclic radicals include,but are not limited to, azepinyl, benzodioxanyl, benzodioxolyl,benzofuranonyl, benzopyranonyl, benzopyranyl, benzotetrahydrofuranyl,benzotetrahydrothienyl, benzothiopyranyl, benzoxazinyl, β-carbolinyl,chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl,dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dihydrofuryl,dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl,dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolanyl,1,4-dithianyl, furanonyl, imidazolidinyl, imidazolinyl, indolinyl,isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isochromanyl,isocoumarinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl,morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinonyl,oxazolidinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidonyl,pyrazolidinyl, pyrazolinyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl,tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydropyranyl,tetrahydrothienyl, thiamorpholinyl, thiazolidinyl, tetrahydroquinolinyl,and 1,3,5-trithianyl. In certain embodiments, heterocyclic may also beoptionally substituted as described herein. Non-limiting examples ofmoieties with which the heterocyclic group can be substituted arehalogen (fluoro, chloro, bromo or iodo), hydroxyl, carbonyl,alkoxycarbonyl, alkoxycarbonylalkyl, sulfanyl, amino, alkylamino,arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate,phosphonic acid, phosphate, and phosphonate, either unprotected, orprotected as necessary.

The term “heteroaryl” refers to refers to a monovalent monocyclicaromatic group and/or multicyclic aromatic group that contain at leastone aromatic ring, wherein at least one aromatic ring contains one ormore heteroatoms independently selected from O, S, and N in the ring.Heteroaryl groups are bonded to the rest of the molecule through thearomatic ring. Each ring of a heteroaryl group can contain one or two Oatoms, one or two S atoms, and/or one to four N atoms, provided that thetotal number of heteroatoms in each ring is four or less and each ringcontains at least one carbon atom. In certain embodiments, theheteroaryl has from 5 to 20, from 5 to 15, or from 5 to 10 ring atoms.Examples of monocyclic heteroaryl groups include, but are not limitedto, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl,oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl,pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl,triazinyl, and triazolyl. Examples of bicyclic heteroaryl groupsinclude, but are not limited to, benzofuranyl, benzimidazolyl,benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl,benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl,imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl,isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl,isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl,pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl,quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl, and thienopyridyl.Examples of tricyclic heteroaryl groups include, but are not limited to,acridinyl, benzindolyl, carbazolyl, dibenzofuranyl, perimidinyl,phenanthrolinyl, phenanthridinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxazinyl, and xanthenyl. In certain embodiments,heteroaryl may also be optionally substituted as described herein.

The term “alkylaryl” refers to an aryl group with an alkyl substituent.The term “aralkyl” or “arylalkyl” refers to an alkyl group with an arylsubstituent.

The term “alkylheterocyclo” refers to a heterocyclo group with an alkylsubstituent. The term “heterocycloalkyl” refers to an alkyl group with aheterocyclo substituent.

The term “alkylheteroaryl” refers to a heteroaryl group with an alkylsubstituent. The term “heteroarylalkyl” refers to an alkyl group with aheteroaryl substituent.

As used herein, the term “hydantoinyl” refers to the group

where R^(XX) and R^(YY) are each independently hydrogen or lower alkyl.

As used herein, the term “hydantoinylalkyl” refers to the group-alkyl-hydantoinyl, where alkyl and hydantoinyl are as described herein.

The term “protecting group” as used herein and unless otherwise definedrefers to a group that is added to an oxygen, nitrogen, or phosphorusatom to prevent its further reaction or for other purposes. A widevariety of oxygen and nitrogen protecting groups are known to thoseskilled in the art of organic synthesis.

“Pharmaceutically acceptable salt” refers to any salt of a compoundprovided herein which retains its biological properties and which is nottoxic or otherwise undesirable for pharmaceutical use. Such salts may bederived from a variety of organic and inorganic counter-ions well knownin the art. Such salts include, but are not limited to: (1) acidaddition salts formed with organic or inorganic acids such ashydrochloric, hydrobromic, sulfuric, nitric, phosphoric, sulfamic,acetic, trifluoroacetic, trichloroacetic, propionic, hexanoic,cyclopentylpropionic, glycolic, glutaric, pyruvic, lactic, malonic,succinic, sorbic, ascorbic, malic, maleic, fumaric, tartaric, citric,benzoic, 3-(4-hydroxybenzoyl)benzoic, picric, cinnamic, mandelic,phthalic, lauric, methanesulfonic, ethanesulfonic,1,2-ethane-disulfonic, 2-hydroxyethanesulfonic, benzenesulfonic,4-chlorobenzenesulfonic, 2-naphthalenesulfonic, 4-toluenesulfonic,camphoric, camphorsulfonic,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic, glucoheptonic,3-phenylpropionic, trimethylacetic, tert-butylacetic, lauryl sulfuric,gluconic, benzoic, glutamic, hydroxynaphthoic, salicylic, stearic,cyclohexylsulfamic, quinic, muconic acid and the like acids; or (2) baseaddition salts formed when an acidic proton present in the parentcompound either (a) is replaced by a metal ion, e.g., an alkali metalion, an alkaline earth ion or an aluminum ion, or alkali metal oralkaline earth metal hydroxides, such as sodium, potassium, calcium,magnesium, aluminum, lithium, zinc, and barium hydroxide, ammonia or (b)coordinates with an organic base, such as aliphatic, alicyclic, oraromatic organic amines, such as ammonia, methylamine, dimethylamine,diethylamine, picoline, ethanolamine, diethanolamine, triethanolamine,ethylenediamine, lysine, arginine, ornithine, choline,N,N′-dibenzylethylene-diamine, chloroprocaine, diethanolamine, procaine,N-benzylphenethylamine, N-methylglucamine piperazine,tris(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, and thelike.

Pharmaceutically acceptable salts further include, by way of exampleonly and without limitation, sodium, potassium, calcium, magnesium,ammonium, tetraalkylammonium and the like, and when the compoundcontains a basic functionality, salts of non-toxic organic or inorganicacids, such as hydrohalides, e.g. hydrochloride and hydrobromide,sulfate, phosphate, sulfamate, nitrate, acetate, trifluoroacetate,trichloroacetate, propionate, hexanoate, cyclopentylpropionate,glycolate, glutarate, pyruvate, lactate, malonate, succinate, sorbate,ascorbate, malate, maleate, fumarate, tartarate, citrate, benzoate,3-(4-hydroxybenzoyl)benzoate, picrate, cinnamate, mandelate, phthalate,laurate, methanesulfonate (mesylate), ethanesulfonate,1,2-ethane-disulfonate, 2-hydroxyethanesulfonate, benzenesulfonate(besylate), 4-chlorobenzenesulfonate, 2-naphthalenesulfonate,4-toluenesulfonate, camphorate, camphorsulfonate,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate, glucoheptonate,3-phenylpropionate, trimethylacetate, tert-butylacetate, lauryl sulfate,gluconate, benzoate, glutamate, hydroxynaphthoate, salicylate, stearate,cyclohexylsulfamate, quinate, muconate and the like.

As used herein, the term “nucleobase” refers to the base portion of anucleoside or nucleotide. In certain embodiments, a nucleobase is apurine or pyrimidine base, as defined herein.

The term “purine” or “pyrimidine” base refers to, but is not limited to,adenine, N⁶-alkylpurines, N⁶-acylpurines (wherein acyl is C(O)(alkyl,aryl, alkylaryl, or arylalkyl), N⁶-benzylpurine, N⁶-halopurine,N⁶-vinylpurine, N⁶-acetylenic purine, N⁶-acyl purine, N⁶-hydroxyalkylpurine, N⁶-alkylaminopurine, N⁶-thioalkyl purine, N²-alkylpurines,N²-alkyl-6-thiopurines, thymine, cytosine, 5-fluorocytosine,5-methylcytosine, 6-azapyrimidine, including 6-azacytosine, 2- and/or4-mercaptopyrmidine, uracil, 5-halouracil, including 5-fluorouracil,C⁵-alkylpyrimidines, C⁵-benzylpyrimidines, C⁵-halopyrimidines,C⁵-vinylpyrimidine, C⁵-acetylenic pyrimidine, C⁵-acyl pyrimidine,C⁵-hydroxyalkyl purine, C⁵-amidopyrimidine, C⁵-cyanopyrimidine,C⁵-iodopyrimidine, C⁶-iodo-pyrimidine, C⁵—Br-vinyl pyrimidine,C⁶—Br-vinyl pyrimidine, C⁵-nitropyrimidine, C⁵-amino-pyrimidine,N²-alkylpurines, N²-alkyl-6-thiopurines, 5-azacytidinyl, 5-azauracilyl,triazolopyridinyl, imidazolopyridinyl, pyrrolopyrimidinyl,pyrrolotriazine, and pyrazolopyrimidinyl. Purine bases include, but arenot limited to, guanine, adenine, hypoxanthine, 7-deazaguanine,7-fluoro-7-deazaguanine, 7-deazaadenine, 7-fluoro-7-deazaadenine,2,6-diaminopurine, 2-amino-6-chloropurine, 6-ethoxypurine,6-methoxylpurine and 6-chloropurine. Functional oxygen and nitrogengroups on the base can be protected as necessary or desired. Suitableprotecting groups are well known to those skilled in the art, andinclude trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, andt-butyldiphenylsilyl, trityl, alkyl groups, and acyl groups such asacetyl and propionyl, methanesulfonyl, and p-toluenesulfonyl.

The term “acyl” or “O-linked ester” refers to a group of the formulaC(O)R′, wherein R′ is alkyl or cycloalkyl (including lower alkyl),carboxylate reside of amino acid, aryl including phenyl, alkaryl,arylalkyl including benzyl, alkoxyalkyl including methoxymethyl,aryloxyalkyl such as phenoxymethyl; or substituted alkyl (includinglower alkyl), aryl including phenyl optionally substituted with chloro,bromo, fluoro, iodo, C₁ to C₄ alkyl or C₁ to C₄ alkoxy, sulfonate esterssuch as alkyl or arylalkyl sulphonyl including methanesulfonyl, themono, di or triphosphate ester, trityl or monomethoxy-trityl,substituted benzyl, alkaryl, arylalkyl including benzyl, alkoxyalkylincluding methoxymethyl, aryloxyalkyl such as phenoxymethyl. Aryl groupsin the esters optimally comprise a phenyl group. In particular, acylgroups include acetyl, trifluoroacetyl, methylacetyl, cyclpropylacetyl,propionyl, butyryl, hexanoyl, heptanoyl, octanoyl, neo-heptanoyl,phenylacetyl, 2-acetoxy-2-phenylacetyl, diphenylacetyl,α-methoxy-α-trifluoromethyl-phenylacetyl, bromoacetyl,2-nitro-benzeneacetyl, 4-chloro-benzeneacetyl,2-chloro-2,2-diphenylacetyl, 2-chloro-2-phenylacetyl, trimethylacetyl,chlorodifluoroacetyl, perfluoroacetyl, fluoroacetyl,bromodifluoroacetyl, methoxyacetyl, 2-thiopheneacetyl,chlorosulfonylacetyl, 3-methoxyphenylacetyl, phenoxyacetyl,tert-butylacetyl, trichloroacetyl, monochloro-acetyl, dichloroacetyl,7H-dodecafluoro-heptanoyl, perfluoro-heptanoyl,7H-dodeca-fluoroheptanoyl, 7-chlorododecafluoro-heptanoyl,7-chloro-dodecafluoro-heptanoyl, 7H-dodecafluoroheptanoyl,7H-dodeca-fluoroheptanoyl, nona-fluoro-3,6-dioxa-heptanoyl,nonafluoro-3,6-dioxaheptanoyl, perfluoroheptanoyl, methoxybenzoyl,methyl 3-amino-5-phenylthiophene-2-carboxyl,3,6-dichloro-2-methoxy-benzoyl, 4-(1,1,2,2-tetrafluoro-ethoxy)-benzoyl,2-bromo-propionyl, omega-aminocapryl, decanoyl, n-pentadecanoyl,stearyl, 3-cyclopentyl-propionyl, 1-benzene-carboxyl, O-acetylmandelyl,pivaloyl acetyl, 1-adamantane-carboxyl, cyclohexane-carboxyl,2,6-pyridinedicarboxyl, cyclopropane-carboxyl, cyclobutane-carboxyl,perfluorocyclohexyl carboxyl, 4-methylbenzoyl, chloromethyl isoxazolylcarbonyl, perfluorocyclohexyl carboxyl, crotonyl,1-methyl-1H-indazole-3-carbonyl, 2-propenyl, isovaleryl,1-pyrrolidinecarbonyl, 4-phenylbenzoyl.

The term “amino acid” refers to naturally occurring and synthetic α, β,γ, or δ amino acids, and includes but is not limited to, amino acidsfound in proteins, i.e. glycine, alanine, valine, leucine, isoleucine,methionine, phenylalanine, tryptophan, proline, serine, threonine,cysteine, tyrosine, asparagine, glutamine, aspartate, glutamate, lysine,arginine and histidine. In certain embodiments, the amino acid is in theL-configuration. In certain embodiments, the amino acid is in theD-configuration. In certain embodiments, the amino acid is provided as asubstituent of a compound described herein, wherein the amino acid is aresidue of alanyl, valinyl, leucinyl, isoleuccinyl, prolinyl,phenylalaninyl, tryptophanyl, methioninyl, glycinyl, serinyl,threoninyl, cysteinyl, tyrosinyl, asparaginyl, glutaminyl, aspartoyl,glutaroyl, lysinyl, argininyl, histidinyl, β-alanyl, β-valinyl,β-leucinyl, β-isoleuccinyl, β-prolinyl, β-phenylalaninyl,β-tryptophanyl, β-methioninyl, β-glycinyl, β-serinyl, β-threoninyl,β-cysteinyl, β-tyrosinyl, β-asparaginyl, β-glutaminyl, β-aspartoyl,β-glutaroyl, β-lysinyl, β-argininyl, or β-histidinyl.

The term “amino acid derivative” refers to a group derivable from anaturally or non-naturally occurring amino acid, as described andexemplified herein. Amino acid derivatives are apparent to those ofskill in the art and include, but are not limited to, ester, aminoalcohol, amino aldehyde, amino lactone, and N-methyl derivatives ofnaturally and non-naturally occurring amino acids. In an embodiment, anamino acid derivative is provided as a substituent of a compounddescribed herein, wherein the substituent is —NR^(X)-G(S_(C))—C(O)-Q¹,wherein Q¹ is —SR^(Y), —NR^(Y)R^(Y) or alkoxyl, R^(Y) is hydrogen oralkyl, S_(C) is a side chain of a naturally occurring or non-naturallyoccurring amino acid, G is C₁-C₂ alkyl, and R^(X) is hydrogen or R^(X)and S_(C), together with the atoms to which they are attached, combineto form a five-membered heterocyclic ring. In an embodiment, an aminoacid derivative is provided as a substituent of a compound describedherein, wherein the substituent is —O—C(O)-G(S_(C))—NH-Q², wherein Q² ishydrogen or alkoxyl, S_(C) is a side chain of a naturally occurring ornon-naturally occurring amino acid and G is C₁-C₂ alkyl. In certainembodiments, Q² and S_(C), together with the atoms to which they areattached, combine to form a five-membered heterocyclic ring. In certainembodiments, G is C₁ alkyl and S_(C) is hydrogen, alkyl, arylalkyl,heterocycloalkyl, carboxylalkyl, heteroarylalkyl, aminoalkyl,hydroxylalkyl, aminoiminoaminoalkyl, aminocarbonylalkyl, sulfanylalkyl,carbamoylalkyl, alkylsulfanylalkyl, or hydroxylarylalkyl. In anembodiment, an amino acid derivative is provided as a substituent of acompound described herein, wherein the amino acid derivative is in theD-configuration. In an embodiment, an amino acid derivative is providedas a substituent of a compound described herein, wherein the amino acidderivative is in the L-configuration.

As used herein, the term “aminoalkyl” refers to an alkyl group with anamino substituent, where alkyl and amino are as described herein.

As used herein, the term “carboxylalkyl” refers to the group-alkyl-C(O)OH, where alkyl is as described herein.

As used herein, the term “aminoiminoaminoalkyl” refers to the group-alkyl-amino-C(NH)-amino, where alkyl and amino are as described herein.

As used herein, the term “aminocarbonylalkyl” refers to the group-alkyl-C(O)-amino, where alkyl and amino are as described herein.

As used herein, the term “sulfanylalkyl” refers to the group -alkyl-SH,where alkyl is as described herein.

As used herein, the term “carbamoylalkyl” refers to the group-alkyl-C(O)-amino, where alkyl and amino are as described herein.

As used herein, the term “alkylsulfanylalkyl” refers to the group-alkyl-S-alkyl, where alkyl is as described herein.

As used herein, the term “hydroxylarylalkyl” refers to the group-alkyl-aryl-OH, where alkyl and aryl are as described herein.

As used herein when referring to a substituent on a sugar ring of anucleoside, the term “alpha” refers to a substituent on the same side ofthe plane of the sugar ring as the 5′ carbon and the term “beta” refersto a substituent on the opposite side of the plane of the sugar ringfrom the 5′ carbon. As shown below, substituent “A” is in the “alpha”position, and substituent “B” is in the “beta” position with respect tothe 5′ carbon:

The term “substantially free of” or “substantially in the absence of”with respect to a nucleoside composition refers to a nucleosidecomposition that includes at least 85 or 90% by weight, in certainembodiments 95%, 98%, 99% or 100% by weight, of the designatedenantiomer of that nucleoside. In certain embodiments of the methods andcompounds provided herein, the compounds are substantially free of anon-designated enantiomer.

Similarly, the term “isolated” with respect to a nucleoside compositionrefers to a nucleoside composition that includes at least 85%, 90%, 95%,98%, or 99% to 100% by weight, of the nucleoside, the remaindercomprising other chemical species or enantiomers.

“Solvate” refers to a compound provided herein or a salt thereof, thatfurther includes a stoichiometric or non-stoichiometric amount ofsolvent bound by non-covalent intermolecular forces. Where the solventis water, the solvate is a hydrate.

“Isotopic composition” refers to the amount of each isotope present fora given atom, and “natural isotopic composition” refers to the naturallyoccurring isotopic composition or abundance for a given atom. Atomscontaining their natural isotopic composition may also be referred toherein as “non-enriched” atoms. Unless otherwise designated, the atomsof the compounds recited herein are meant to represent any stableisotope of that atom. For example, unless otherwise stated, when aposition is designated specifically as “H” or “hydrogen,” the positionis understood to have hydrogen at its natural isotopic composition.

“Isotopic enrichment” refers to the percentage of incorporation of anamount of a specific isotope at a given atom in a molecule in the placeof that atom's natural isotopic abundance. For example, deuteriumenrichment of 1% at a given position means that 1% of the molecules in agiven sample contain deuterium at the specified position. Because thenaturally occurring distribution of deuterium is about 0.0156%,deuterium enrichment at any position in a compound synthesized usingnon-enriched starting materials is about 0.0156%. The isotopicenrichment of the compounds provided herein can be determined usingconventional analytical methods known to one of ordinary skill in theart, including mass spectrometry and nuclear magnetic resonancespectroscopy.

“Isotopically enriched” refers to an atom having an isotopic compositionother than the natural isotopic composition of that atom. “Isotopicallyenriched” may also refer to a compound containing at least one atomhaving an isotopic composition other than the natural isotopiccomposition of that atom.

As used herein, “alkyl,” “cycloalkyl,” “alkenyl,” “cycloalkenyl,”“alkynyl,” “aryl,” “alkoxy,” “alkoxycarbonyl,” “alkoxycarbonylalkyl,”“amino,” “carboxyl,” “alkylamino,” “arylamino,” “thioalkyoxy,”“heterocyclo,” “heteroaryl,” “alkylheterocyclo,” “alkylheteroaryl,”“acyl,” “aralkyl,” “alkaryl,” “purine,” “pyrimidine,” “carboxyl,” and“amino acid” groups optionally comprise deuterium at one or morepositions where hydrogen atoms are present, and wherein the deuteriumcomposition of the atom or atoms is other than the natural isotopiccomposition.

Also as used herein, “alkyl,” “cyclo alkyl,” “alkenyl,” “cycloalkenyl,”“alkynyl,” “aryl,” “alkoxy,” “alkoxycarbonyl,” “alkoxycarbonylalkyl,”“carboxyl,” “alkylamino,” “arylamino,” “thioalkyoxy,” “heterocyclo,”“heteroaryl,” “alkylheterocyclo,” “alkylheteroaryl,” “acyl,” “aralkyl,”“alkaryl,” “purine,” “pyrimidine,” “carboxyl,” and “amino acid” groupsoptionally comprise carbon-13 at an amount other than the naturalisotopic composition.

As used herein, EC₅₀ refers to a dosage, concentration, or amount of aparticular test compound that elicits a dose-dependent response at 50%of maximal expression of a particular response that is induced,provoked, or potentiated by the particular test compound.

As used herein, the IC₅₀ refers to an amount, concentration, or dosageof a particular test compound that achieves a 50% inhibition of amaximal response in an assay that measures such response.

The term “host,” as used herein, refers to any unicellular ormulticellular organism in which the virus can replicate, including celllines and animals, and in certain embodiments, a human. Alternatively,the host can be carrying a part of the Flaviviridae viral genome, whosereplication or function can be altered by the compounds of the presentinvention. The term host specifically includes infected cells, cellstransfected with all or part of the Flaviviridae genome and animals, inparticular, primates (including chimpanzees) and humans. In most animalapplications of the present invention, the host is a human patient.Veterinary applications, in certain indications, however, are clearlyanticipated by the present invention (such as chimpanzees).

As used herein, the terms “subject” and “patient” are usedinterchangeably herein. The terms “subject” and “subjects” refer to ananimal, such as a mammal including a non-primate (e.g., a cow, pig,horse, cat, dog, rat, and mouse) and a primate (e.g., a monkey such as acynomolgous monkey, a chimpanzee and a human), and for example, a human.In certain embodiments, the subject is refractory or non-responsive tocurrent treatments for hepatitis C infection. In another embodiment, thesubject is a farm animal (e.g., a horse, a cow, a pig, etc.) or a pet(e.g., a dog or a cat). In certain embodiments, the subject is a human.

As used herein, the terms “therapeutic agent” and “therapeutic agents”refer to any agent(s) which can be used in the treatment or preventionof a disorder or one or more symptoms thereof. In certain embodiments,the term “therapeutic agent” includes a compound provided herein. Incertain embodiments, a therapeutic agent is an agent which is known tobe useful for, or has been or is currently being used for the treatmentor prevention of a disorder or one or more symptoms thereof.

“Therapeutically effective amount” refers to an amount of a compound orcomposition that, when administered to a subject for treating a disease,is sufficient to effect such treatment for the disease. A“therapeutically effective amount” can vary depending on, inter alia,the compound, the disease and its severity, and the age, weight, etc.,of the subject to be treated.

“Treating” or “treatment” of any disease or disorder refers, in certainembodiments, to ameliorating a disease or disorder that exists in asubject. In another embodiment, “treating” or “treatment” includesameliorating at least one physical parameter, which may be indiscernibleby the subject. In yet another embodiment, “treating” or “treatment”includes modulating the disease or disorder, either physically (e.g.,stabilization of a discernible symptom) or physiologically (e.g.,stabilization of a physical parameter) or both. In yet anotherembodiment, “treating” or “treatment” includes delaying the onset of thedisease or disorder.

As used herein, the terms “prophylactic agent” and “prophylactic agents”as used refer to any agent(s) which can be used in the prevention of adisorder or one or more symptoms thereof. In certain embodiments, theterm “prophylactic agent” includes a compound provided herein. Incertain other embodiments, the term “prophylactic agent” does not refera compound provided herein. For example, a prophylactic agent is anagent which is known to be useful for, or has been or is currently beingused to prevent or impede the onset, development, progression, and/orseverity of a disorder.

As used herein, the phrase “prophylactically effective amount” refers tothe amount of a therapy (e.g., prophylactic agent) which is sufficientto result in the prevention or reduction of the development, recurrenceor onset of one or more symptoms associated with a disorder (or toenhance or improve the prophylactic effect(s) of another therapy (e.g.,another prophylactic agent).

Compounds

Provided herein are thiophosphate nucleoside compounds useful for thetreatment of Flaviviridae infections such as HCV infection. Thethiophosphate nucleoside compounds can be formed as described herein andused for the treatment of Flaviviridae infections such as HCV infection.

In certain embodiments, provided herein are compounds according toFormula 2001:

-   -   or a pharmaceutically acceptable salt, solvate, stereoisomeric        form, tautomeric form, or polymorphic form thereof, wherein:        -   Base is a nucleobase;        -   R^(A) is hydroxyl or halo;        -   R^(B) is methyl or halo;        -   PD is

-   -   -   X is alkyl, alkoxycarbonylalkyl,            (alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,            heterocycloalkyl, hydantoinylalkyl, or aryl;        -   Y is —NR¹R² or an N-linked or O-linked amino acid residue,            or a derivative thereof;        -   Z is alkyl, alkoxycarbonyl, or alkoxyalkylcarbonyl;        -   A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;        -   R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl,            arylalkyl, or heteroarylalkyl;        -   R² is hydrogen or alkyl;        -   R is hydrogen or alkyl; and        -   each of R¹⁰, R¹¹, and R¹² is independently hydrogen or            alkyl; or R¹⁰ and R¹¹, together with the carbon atoms to            which they are attached, combine to form a five-, six-, or            seven-membered ring.

In certain embodiments, provided herein are compounds according toFormula (I):

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein: Base is anucleobase; R^(A) is hydroxyl or halo; R^(B) is methyl or halo; PD is

X is alkyl, alkoxycarbonylalkyl,(alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,heterocycloalkyl, hydantoinylalkyl, or aryl; Y is —NR¹R² or an N-linkedamino acid, or ester thereof; Z is alkyl, alkoxycarbonyl oralkoxyalkylcarbonyl; A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl, orheteroarylalkyl; R² is hydrogen or alkyl; R is hydrogen or alkyl; andeach of R¹⁰, R¹¹, and R¹² is independently hydrogen or alkyl; or R¹⁰ andR¹¹, together with the carbon atoms to which they are attached, combineto form a five-, six-, or seven-membered ring.

In certain embodiments, provided herein are compounds according toFormula III:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, A, Z, R^(A),R^(B) and R are as described in the context of Formula I or Formula2001.

In certain embodiments, provided herein are compounds according toFormula II:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, X, Y, R^(A)and R^(B) are as described in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according to anyof Formulas IV-VI:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, Z, R^(A) andR^(B) are as described in the context of Formula I. In an embodiment, acompound of any of Formulas 2001, I, III, or IV-VI is provided wherein Zis alkyl. In an embodiment, a compound of any of Formulas 2001, I, III,or IV-VI is provided wherein Z is alkoxycarbonyl. In an embodiment, acompound of any of Formulas 2001, I, III, or IV-VI is provided wherein Zis alkoxyalkylcarbonyl.

In certain embodiments, provided herein are compounds according toFormula XXIX:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; wherein R^(M) is alkyl;and where Base, X, R^(A) and R^(B) are as described in the context ofFormula I or Formula 2001. In an embodiment, a compound of Formula XXIXis provided wherein R^(M) is lower alkyl. In an embodiment, a compoundof Formula XXIX is provided wherein R^(M) is C₁-C₃ alkyl.

In certain embodiments, provided herein are compounds according toFormula VII or VIII:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, X, R^(A) andR^(B) are as described in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according to anyof Formulas (IX)-(XII):

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; wherein each R⁶ isindependently hydrogen, halogen, or alkyl; and each R⁷ is independentlyhydrogen or —NH₂; and where PD, R^(A), and R^(B) are as described in thecontext of Formula I. In an embodiment, each R⁷ is independentlyhydrogen or amino.

In certain embodiments, provided herein are compounds according to anyof Formulas XIII-XVII:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where PD, R^(A), and R^(B)are as described in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according to anyof Formulas XVIII or XIX:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; wherein each of R⁸ and R⁹are independently hydrogen or lower alkyl; and where Base, Y, R^(A), andR^(B) are as described in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according toFormula XXVII:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; wherein Q is —SR²⁰;—NR²¹R²² or —OR²⁰; each of R²⁰, R²¹ and R²² is independently hydrogen oralkyl; and G is C₁-C₂ alkyl; and where Base, Y, R^(A), and R^(B) are asdescribed in the context of Formula I or Formula 2001. In an embodiment,a compound of Formula XXVII is provided wherein G is C₂ alkyl. In anembodiment, a compound of Formula XXVII is provided wherein G issubstituted with —NHC(O)Q. In an embodiment, a compound of Formula XXVIIis provided wherein G is —CH₂CH(NHC(O)Q)-.

In certain embodiments, provided herein are compounds according toFormula) XXVIII:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; wherein R^(N) iscycloalkyl or aralkyl; and where Base, A, R, R^(A), and R^(B) are asdescribed in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according to anyof Formulas 2001, I-XXII, XXVII or XXVIII wherein R^(A) is hydroxyl andR^(B) is lower alkyl. In certain embodiments, provided herein arecompounds according to any of Formulas 2001, I-XXII, XXVII or XXVIII,wherein R^(A) is halo and R^(B) is lower alkyl. In certain embodiments,provided herein are compounds according to any of Formulas 2001, I-XXII,XXVII or XXVIII, wherein R^(A) is fluoro and R^(B) is lower alkyl. Incertain embodiments, provided herein are compounds according to any ofFormulas 2001, I-XXII, XXVII or XXVIII, wherein R^(A) is chloro andR^(B) is lower alkyl.

In certain embodiments, provided herein are compounds according to anyof Formulas XX-XXII:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, PD, and R^(B)are as described in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according to anyof Formulas XXIII-XXV:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base and PD are asdescribed in the context of Formula I or Formula 2001.

In certain embodiments, provided herein are compounds according toFormula XXVI:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof; where Base, R^(A) and PDare as described in the context of Formula I or Formula 2001.

In certain embodiments provided herein are compounds according to any ofFormulas I, IX-XVII and XX-XXVI wherein each PD is independently

where X and Y are as described in the context of Formula I. In certainembodiments provided herein are compounds according to any of FormulasI, IX-XVII and XX-XXVI wherein each PD is independently

where Z, R and A are as described in the context of Formula I or Formula2001.

In certain embodiments, provided herein are compounds according to anyof Formulas I-VIII or XVIII-XXIX wherein each Base is independently:

or tautomeric form thereof, wherein: each R⁴ is independently hydrogen,hydroxyl, alkoxyl, amino or aminoalkyl; each R⁵ is independentlyhydrogen, hydroxyl, amino, or alkoxyl; each R⁶ is independentlyhydrogen, halogen, or alkyl; and each R⁷ is independently hydrogen or—NH₂. In an embodiment, each R⁷ is independently hydrogen or amino.

In certain embodiments provided herein is a compound according to any offormulas 2001, I, II, VII-XVII, XX-XXVI or XXIX wherein: X is

n is an integer selected over the range of 1-10; A^(A) is —C(O)—O—R¹⁰⁰;A^(N) is —NH—C(O)—O—R¹⁰¹; and each of R¹⁰⁰ and R¹⁰¹ is independentlylower alkyl. In an embodiment, each of R¹⁰⁰ and R¹⁰¹ is independentlyC₁-C₅ alkyl.

In certain embodiments provided herein are compounds according to any ofFormulas 1-23byii:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof.

In certain embodiments, provided herein are compounds according toFormula (1001):

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein: R^(A) is hydroxylor halo; R^(B) is methyl or halo; PD is

X is alkyl, alkoxycarbonylalkyl,(alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,heterocycloalkyl, hydantoinylalkyl, or aryl; Y is —NR¹R² or an N-linkedamino acid, or ester thereof; Z is alkyl, alkoxycarbonyl oralkoxyalkylcarbonyl; A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl, orheteroarylalkyl; R² is hydrogen or alkyl; R is hydrogen or alkyl; andeach of R¹⁰, R¹¹, and R¹² is independently hydrogen or alkyl; or R¹⁰ andR¹¹, together with the carbon atoms to which they are attached, combineto form a five-, six-, or seven-membered ring.

In certain embodiments, provided herein are compounds according toFormula 1002:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein: R^(A) is hydroxylor halo; and R^(B) is methyl or halo.

In some embodiments, provided herein are:

-   (a) compounds as described herein, e.g., of Formula 2001, 2001,    I-XXIX, 1-23byii and 1001-1002, and pharmaceutically acceptable    salts and compositions thereof;-   (b) compounds as described herein, e.g., of Formula 2001, I-XXIX,    1-23byii and 1001-1002, and pharmaceutically acceptable salts and    compositions thereof for use in the treatment and/or prophylaxis of    a liver disorder including Flaviviridae infection, especially in    individuals diagnosed as having a Flaviviridae infection or being at    risk of becoming infected by hepatitis C;-   (c) processes for the preparation of compounds as described herein,    e.g., of Formula 2001, I-XXIX, 1-23byii and 1001-1002, as described    in more detail elsewhere herein;-   (d) pharmaceutical formulations comprising a compound as described    herein, e.g., of Formula 2001, I-XXIX, 1-23byii and 1001-1002, or a    pharmaceutically acceptable salt thereof together with a    pharmaceutically acceptable carrier or diluent;-   (e) pharmaceutical formulations comprising a compound as described    herein, e.g., of Formula 2001, I-XXIX, 1-23byii and 1001-1002, or a    pharmaceutically acceptable salt thereof together with one or more    other effective anti-HCV agents, optionally in a pharmaceutically    acceptable carrier or diluent;-   (f) a method for the treatment and/or prophylaxis of a host infected    with Flaviviridae that includes the administration of an effective    amount of a compound as described herein, e.g., of Formula 2001,    I-XXIX, 1-23byii and 1001-1002, its pharmaceutically acceptable salt    or composition; or-   (g) a method for the treatment and/or prophylaxis of a host infected    with Flaviviridae that includes the administration of an effective    amount of a compounds as described herein, e.g., of Formula 2001,    I-XXIX, 1-23byii and 1001-1002, its pharmaceutically acceptable salt    or composition in combination and/or alternation with one or more    effective anti-HCV agent.    Optically Active Compounds

It is appreciated that compounds provided herein have several chiralcenters and may exist in and be isolated in optically active and racemicforms. Some compounds may exhibit polymorphism. It is to be understoodthat any racemic, optically-active, diastereomeric, polymorphic, orstereoisomeric form, or mixtures thereof, of a compound provided herein,which possess the useful properties described herein is within the scopeof the invention. It being well known in the art how to prepareoptically active forms (for example, by resolution of the racemic formby recrystallization techniques, by synthesis from optically-activestarting materials, by chiral synthesis, or by chromatographicseparation using a chiral stationary phase).

In particular, since the 1′ and 4′ carbons of a nucleoside are chiral,their non-hydrogen substituents (the base and the CHOR groups,respectively) can be either cis (on the same side) or trans (on oppositesides) with respect to the sugar ring system. The four optical isomerstherefore are represented by the following configurations (whenorienting the sugar moiety in a horizontal plane such that the oxygenatom is in the back): cis (with both groups “up”, which corresponds tothe configuration of naturally occurring β-D nucleosides), cis (withboth groups “down”, which is a non-naturally occurring β-Lconfiguration), trans (with the C2′ substituent “up” and the C4′substituent “down”), and trans (with the C2′ substituent “down” and theC4′ substituent “up”). The “D-nucleosides” are cis nucleosides in anatural configuration and the “L-nucleosides” are cis nucleosides in thenon-naturally occurring configuration.

Likewise, most amino acids are chiral (designated as L or D, wherein theL enantiomer is the naturally occurring configuration) and can exist asseparate enantiomers.

Examples of methods to obtain optically active materials are known inthe art, and include at least the following.

-   -   i) physical separation of crystals—a technique whereby        macroscopic crystals of the individual enantiomers are manually        separated. This technique can be used if crystals of the        separate enantiomers exist, i.e., the material is a        conglomerate, and the crystals are visually distinct;    -   ii) simultaneous crystallization—a technique whereby the        individual enantiomers are separately crystallized from a        solution of the racemate, possible only if the latter is a        conglomerate in the solid state;    -   iii) enzymatic resolutions—a technique whereby partial or        complete separation of a racemate by virtue of differing rates        of reaction for the enantiomers with an enzyme;    -   iv) enzymatic asymmetric synthesis—a synthetic technique whereby        at least one step of the synthesis uses an enzymatic reaction to        obtain an enantiomerically pure or enriched synthetic precursor        of the desired enantiomer;    -   v) chemical asymmetric synthesis—a synthetic technique whereby        the desired enantiomer is synthesized from an achiral precursor        under conditions that produce asymmetry (i.e., chirality) in the        product, which may be achieved using chiral catalysts or chiral        auxiliaries;    -   vi) diastereomer separations—a technique whereby a racemic        compound is reacted with an enantiomerically pure reagent (the        chiral auxiliary) that converts the individual enantiomers to        diastereomers. The resulting diastereomers are then separated by        chromatography or crystallization by virtue of their now more        distinct structural differences and the chiral auxiliary later        removed to obtain the desired enantiomer;    -   vii) first- and second-order asymmetric transformations—a        technique whereby diastereomers from the racemate equilibrate to        yield a preponderance in solution of the diastereomer from the        desired enantiomer or where preferential crystallization of the        diastereomer from the desired enantiomer perturbs the        equilibrium such that eventually in principle all the material        is converted to the crystalline diastereomer from the desired        enantiomer. The desired enantiomer is then released from the        diastereomer;    -   viii) kinetic resolutions—this technique refers to the        achievement of partial or complete resolution of a racemate (or        of a further resolution of a partially resolved compound) by        virtue of unequal reaction rates of the enantiomers with a        chiral, non-racemic reagent or catalyst under kinetic        conditions;    -   ix) enantiospecific synthesis from non-racemic precursors—a        synthetic technique whereby the desired enantiomer is obtained        from non-chiral starting materials and where the stereochemical        integrity is not or is only minimally compromised over the        course of the synthesis;    -   x) chiral liquid chromatography—a technique whereby the        enantiomers of a racemate are separated in a liquid mobile phase        by virtue of their differing interactions with a stationary        phase. The stationary phase can be made of chiral material or        the mobile phase can contain an additional chiral material to        provoke the differing interactions;    -   xi) chiral gas chromatography—a technique whereby the racemate        is volatilized and enantiomers are separated by virtue of their        differing interactions in the gaseous mobile phase with a column        containing a fixed non-racemic chiral adsorbent phase;    -   xii) extraction with chiral solvents—a technique whereby the        enantiomers are separated by virtue of preferential dissolution        of one enantiomer into a particular chiral solvent;    -   xiii) transport across chiral membranes—a technique whereby a        racemate is placed in contact with a thin membrane barrier. The        barrier typically separates two miscible fluids, one containing        the racemate, and a driving force such as concentration or        pressure differential causes preferential transport across the        membrane barrier. Separation occurs as a result of the        non-racemic chiral nature of the membrane which allows only one        enantiomer of the racemate to pass through.

In some embodiments, provided is a composition of a thiophosphatenucleoside compound that comprises a substantially pure designatedenantiomer of the thiophosphate nucleoside compound. In certainembodiments, in the methods and compounds of this invention, thecompounds are substantially free of other enantiomers. In someembodiments, a composition includes a compound that is at least 85%,90%, 95%, 98%, 99%, or 100% by weight, of the compound, the remaindercomprising other chemical species or enantiomers.

Isotopically Enriched Compounds

Also provided herein are isotopically enriched compounds, including butnot limited to isotopically enriched thiophosphate nucleoside compounds.

Isotopic enrichment (for example, deuteration) of pharmaceuticals toimprove pharmacokinetics (“PK”), pharmacodynamics (“PD”), and toxicityprofiles, has been demonstrated previously with some classes of drugs.See, for example, Lijinsky et. al., Food Cosmet. Toxicol., 20: 393(1982); Lijinsky et. al., J. Nat. Cancer Inst., 69: 1127 (1982); Mangoldet. al., Mutation Res. 308: 33 (1994); Gordon et. al., Drug Metab.Dispos., 15: 589 (1987); Zello et. al., Metabolism, 43: 487 (1994);Gately et. al., J. Nucl. Med., 27: 388 (1986); Wade D, Chem. Biol.Interact. 117: 191 (1999).

Isotopic enrichment of a drug can be used, for example, to (1) reduce oreliminate unwanted metabolites, (2) increase the half-life of the parentdrug, (3) decrease the number of doses needed to achieve a desiredeffect, (4) decrease the amount of a dose necessary to achieve a desiredeffect, (5) increase the formation of active metabolites, if any areformed, and/or (6) decrees the production of deleterious metabolites inspecific tissues and/or create a more effective drug and/or a safer drugfor combination therapy, whether the combination therapy is intentionalor not.

Replacement of an atom for one of its isotopes often will result in achange in the reaction rate of a chemical reaction. This phenomenon isknown as the Kinetic Isotope Effect (“KIE”). For example, if a C—H bondis broken during a rate-determining step in a chemical reaction (i.e.the step with the highest transition state energy), substitution of adeuterium for that hydrogen will cause a decrease in the reaction rateand the process will slow down. This phenomenon is known as theDeuterium Kinetic Isotope Effect (“DKIE”). (See, e.g., Foster et al.,Adv. Drug Res., vol. 14, pp. 1-36 (1985); Kushner et al., Can. J.Physiol. Pharmacol., vol. 77, pp. 79-88 (1999)).

The magnitude of the DKIE can be expressed as the ratio between therates of a given reaction in which a C—H bond is broken, and the samereaction where deuterium is substituted for hydrogen. The DKIE can rangefrom about 1 (no isotope effect) to very large numbers, such as 50 ormore, meaning that the reaction can be fifty, or more, times slower whendeuterium is substituted for hydrogen. High DKIE values may be due inpart to a phenomenon known as tunneling, which is a consequence of theuncertainty principle. Tunneling is ascribed to the small mass of ahydrogen atom, and occurs because transition states involving a protoncan sometimes form in the absence of the required activation energy.Because deuterium has more mass than hydrogen, it statistically has amuch lower probability of undergoing this phenomenon.

Tritium (“T”) is a radioactive isotope of hydrogen, used in research,fusion reactors, neutron generators, and radiopharmaceuticals. Tritiumis a hydrogen atom that has 2 neutrons in the nucleus and has an atomicweight close to 3. It occurs naturally in the environment in very lowconcentrations, most commonly found as T₂O. Tritium decays slowly(half-life=12.3 years) and emits a low energy beta particle that cannotpenetrate the outer layer of human skin. Internal exposure is the mainhazard associated with this isotope, yet it must be ingested in largeamounts to pose a significant health risk. As compared with deuterium, alesser amount of tritium must be consumed before it reaches a hazardouslevel. Substitution of tritium (“T”) for hydrogen results in yet astronger bond than deuterium and gives numerically larger isotopeeffects. Similarly, substitution of isotopes for other elements,including, but not limited to, ¹³C or ¹⁴C for carbon, ³³S, ³⁴S, or ³⁶Sfor sulfur, ¹⁵N for nitrogen, and ¹⁷O or ¹⁸O for oxygen, may lead to asimilar kinetic isotope effect.

For example, the DKIE was used to decrease the hepatotoxicity ofhalothane by presumably limiting the production of reactive species suchas trifluoroacetyl chloride. However, this method may not be applicableto all drug classes. For example, deuterium incorporation can lead tometabolic switching. The concept of metabolic switching asserts thatxenogens, when sequestered by Phase I enzymes, may bind transiently andre-bind in a variety of conformations prior to the chemical reaction(e.g., oxidation). This hypothesis is supported by the relatively vastsize of binding pockets in many Phase I enzymes and the promiscuousnature of many metabolic reactions. Metabolic switching can potentiallylead to different proportions of known metabolites as well as altogethernew metabolites. This new metabolic profile may impart more or lesstoxicity.

The animal body expresses a variety of enzymes for the purpose ofeliminating foreign substances, such as therapeutic agents, from itscirculation system. Examples of such enzymes include the cytochrome P450enzymes (“CYPs”), esterases, proteases, reductases, dehydrogenases, andmonoamine oxidases, to react with and convert these foreign substancesto more polar intermediates or metabolites for renal excretion. Some ofthe most common metabolic reactions of pharmaceutical compounds involvethe oxidation of a carbon-hydrogen (C—H) bond to either a carbon-oxygen(C—O) or carbon-carbon (C—C) pi-bond. The resultant metabolites may bestable or unstable under physiological conditions, and can havesubstantially different pharmacokinetic, pharmacodynamic, and acute andlong-term toxicity profiles relative to the parent compounds. For manydrugs, such oxidations are rapid. These drugs therefore often requirethe administration of multiple or high daily doses.

Therefore, isotopic enrichment at certain positions of a compoundprovided herein will produce a detectable KIE that will affect thepharmacokinetic, pharmacologic, and/or toxicological profiles of acompound provided herein in comparison with a similar compound having anatural isotopic composition.

Preparation of Compounds

The compounds provided herein can be prepared, isolated, or obtained byany method apparent to those of skill in the art. Compounds providedherein can be prepared according to the Exemplary Preparation Schemeprovided below. Reaction conditions, steps, and reactants not providedin the Exemplary Preparation Scheme would be apparent to, and known by,those skilled in the art.

Exemplary Preparation Schemes

In the Exemplary Preparation Schemes, Base, R^(A), R^(B), X, Y, Z and Aare as described in the context of Formula I or 2001. Additional stepsand reagents not provided in the Exemplary Preparation Schemes would beknown to those of skill in the art. Exemplary methods of preparation aredescribed in detail in the Examples herein.

Pharmaceutical Compositions and Methods of Administration

Thiophosphate nucleoside compounds can be formulated into pharmaceuticalcompositions using methods available in the art and those disclosedherein. Any of the compounds disclosed herein can be provided in theappropriate pharmaceutical composition and be administered by a suitableroute of administration.

The methods provided herein encompass administering pharmaceuticalcompositions containing at least one compound as described herein,including a compound of general Formula 2001, I-XXIX, 1-23byii and1001-1002, if appropriate in the salt form, either used alone or in theform of a combination with one or more compatible and pharmaceuticallyacceptable carriers, such as diluents or adjuvants, or with anotheranti-HCV agent.

In certain embodiments, the second agent can be formulated or packagedwith the compound provided herein. Of course, the second agent will onlybe formulated with the compound provided herein when, according to thejudgment of those of skill in the art, such co-formulation should notinterfere with the activity of either agent or the method ofadministration. In certain embodiments, the compound provided herein andthe second agent are formulated separately. They can be packagedtogether, or packaged separately, for the convenience of thepractitioner of skill in the art.

In clinical practice the active agents provided herein may beadministered by any conventional route, in particular orally,parenterally, rectally or by inhalation (e.g. in the form of aerosols).In certain embodiments, the compound provided herein is administeredorally.

Use may be made, as solid compositions for oral administration, oftablets, pills, hard gelatin capsules, powders or granules. In thesecompositions, the active product is mixed with one or more inertdiluents or adjuvants, such as sucrose, lactose, or starch.

These compositions can comprise substances other than diluents, forexample a lubricant, such as magnesium stearate, or a coating intendedfor controlled release.

Use may be made, as liquid compositions for oral administration, ofsolutions which are pharmaceutically acceptable, suspensions, emulsions,syrups, and elixirs containing inert diluents, such as water or liquidparaffin. These compositions can also comprise substances other thandiluents, for example wetting, sweetening, or flavoring products.

The compositions for parenteral administration can be emulsions orsterile solutions. Use may be made, as solvent or vehicle, of propyleneglycol, a polyethylene glycol, vegetable oils, in particular olive oil,or injectable organic esters, for example ethyl oleate. Thesecompositions can also contain adjuvants, in particular wetting,isotonizing, emulsifying, dispersing, and stabilizing agents.Sterilization can be carried out in several ways, for example using abacteriological filter, by radiation or by heating. They can also beprepared in the form of sterile solid compositions which can bedissolved at the time of use in sterile water or any other injectablesterile medium.

The compositions for rectal administration are suppositories or rectalcapsules which contain, in addition to the active principle, excipientssuch as cocoa butter, semi-synthetic glycerides, or polyethyleneglycols.

The compositions can also be aerosols. For use in the form of liquidaerosols, the compositions can be stable sterile solutions or solidcompositions dissolved at the time of use in apyrogenic sterile water,in saline or any other pharmaceutically acceptable vehicle. For use inthe form of dry aerosols intended to be directly inhaled, the activeprinciple is finely divided and combined with a water-soluble soliddiluent or vehicle, for example dextran, mannitol or lactose.

In certain embodiments, a composition provided herein is apharmaceutical composition or a single unit dosage form. Pharmaceuticalcompositions and single unit dosage forms provided herein comprise aprophylactically or therapeutically effective amount of one or moreprophylactic or therapeutic agents (e.g., a compound provided herein, orother prophylactic or therapeutic agent), and a typically one or morepharmaceutically acceptable carriers or excipients. In a specificembodiment and in this context, the term “pharmaceutically acceptable”means approved by a regulatory agency of the Federal or a stategovernment or listed in the U.S. Pharmacopeia or other generallyrecognized pharmacopeia for use in animals, and more particularly inhumans. The term “carrier” includes a diluent, adjuvant (e.g., Freund'sadjuvant (complete and incomplete)), excipient, or vehicle with whichthe therapeutic is administered. Such pharmaceutical carriers can besterile liquids, such as water and oils, including those of petroleum,animal, vegetable or synthetic origin, such as peanut oil, soybean oil,mineral oil, sesame oil and the like. Water can be used as a carrierwhen the pharmaceutical composition is administered intravenously.Saline solutions and aqueous dextrose and glycerol solutions can also beemployed as liquid carriers, particularly for injectable solutions.Examples of suitable pharmaceutical carriers are described in“Remington's Pharmaceutical Sciences” by E. W. Martin.

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well-known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsinclude starch, glucose, lactose, sucrose, gelatin, malt, rice, flour,chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodiumchloride, dried skim milk, glycerol, propylene, glycol, water, ethanol,and the like. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a subjectand the specific active ingredients in the dosage form. The compositionor single unit dosage form, if desired, can also contain minor amountsof wetting or emulsifying agents, or pH buffering agents.

Lactose free compositions provided herein can comprise excipients thatare well known in the art and are listed, for example, in the U.S.Pharmocopia (USP) SP (XXI)/NF (XVI). In general, lactose freecompositions comprise an active ingredient, a binder/filler, and alubricant in pharmaceutically compatible and pharmaceutically acceptableamounts. Exemplary lactose free dosage forms comprise an activeingredient, microcrystalline cellulose, pre gelatinized starch, andmagnesium stearate.

Further encompassed herein are anhydrous pharmaceutical compositions anddosage forms comprising active ingredients, since water can facilitatethe degradation of some compounds. For example, the addition of water(e.g., 5%) is widely accepted in the pharmaceutical arts as a means ofsimulating long term storage in order to determine characteristics suchas shelf life or the stability of formulations over time. See, e.g.,Jens T. Carstensen, Drug Stability: Principles & Practice, 2d. Ed.,Marcel Dekker, New York, 1995, pp. 379 80. In effect, water and heataccelerate the decomposition of some compounds. Thus, the effect ofwater on a formulation can be of great significance since moistureand/or humidity are commonly encountered during manufacture, handling,packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms provided hereincan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine can be anhydrousif substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions can be packaged using materials known to prevent exposureto water such that they can be included in suitable formulary kits.Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

Further provided are pharmaceutical compositions and dosage forms thatcomprise one or more compounds that reduce the rate by which an activeingredient will decompose. Such compounds, which are referred to hereinas “stabilizers,” include, but are not limited to, antioxidants such asascorbic acid, pH buffers, or salt buffers.

The pharmaceutical compositions and single unit dosage forms can takethe form of solutions, suspensions, emulsion, tablets, pills, capsules,powders, sustained-release formulations and the like. Oral formulationcan include standard carriers such as pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, sodium saccharine, cellulose,magnesium carbonate, etc. Such compositions and dosage forms willcontain a prophylactically or therapeutically effective amount of aprophylactic or therapeutic agent, in certain embodiments, in purifiedform, together with a suitable amount of carrier so as to provide theform for proper administration to the subject. The formulation shouldsuit the mode of administration. In a certain embodiment, thepharmaceutical compositions or single unit dosage forms are sterile andin suitable form for administration to a subject, for example, an animalsubject, such as a mammalian subject, for example, a human subject.

A pharmaceutical composition is formulated to be compatible with itsintended route of administration. Examples of routes of administrationinclude, but are not limited to, parenteral, e.g., intravenous,intradermal, subcutaneous, intramuscular, subcutaneous, oral, buccal,sublingual, inhalation, intranasal, transdermal, topical, transmucosal,intra-tumoral, intra-synovial, and rectal administration. In a specificembodiment, the composition is formulated in accordance with routineprocedures as a pharmaceutical composition adapted for intravenous,subcutaneous, intramuscular, oral, intranasal, or topical administrationto human beings. In an embodiment, a pharmaceutical composition isformulated in accordance with routine procedures for subcutaneousadministration to human beings. Typically, compositions for intravenousadministration are solutions in sterile isotonic aqueous buffer. Wherenecessary, the composition may also include a solubilizing agent and alocal anesthetic such as lignocamne to ease pain at the site of theinjection.

Examples of dosage forms include, but are not limited to: tablets;caplets; capsules, such as soft elastic gelatin capsules; cachets;troches; lozenges; dispersions; suppositories; ointments; cataplasms(poultices); pastes; powders; dressings; creams; plasters; solutions;patches; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosageforms suitable for oral or mucosal administration to a subject,including suspensions (e.g., aqueous or non-aqueous liquid suspensions,oil in water emulsions, or a water in oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a subject; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a subject.

The composition, shape, and type of dosage forms provided herein willtypically vary depending on their use. For example, a dosage form usedin the initial treatment of viral infection may contain larger amountsof one or more of the active ingredients it comprises than a dosage formused in the maintenance treatment of the same infection. Similarly, aparenteral dosage form may contain smaller amounts of one or more of theactive ingredients it comprises than an oral dosage form used to treatthe same disease or disorder. These and other ways in which specificdosage forms encompassed herein will vary from one another will bereadily apparent to those skilled in the art. See, e.g., Remington'sPharmaceutical Sciences, 20th ed., Mack Publishing, Easton Pa. (2000).

Generally, the ingredients of compositions are supplied eitherseparately or mixed together in unit dosage form, for example, as a drylyophilized powder or water free concentrate in a hermetically sealedcontainer such as an ampoule or sachette indicating the quantity ofactive agent. Where the composition is to be administered by infusion,it can be dispensed with an infusion bottle containing sterilepharmaceutical grade water or saline. Where the composition isadministered by injection, an ampoule of sterile water for injection orsaline can be provided so that the ingredients may be mixed prior toadministration.

Typical dosage forms comprise a compound provided herein, or apharmaceutically acceptable salt, solvate or hydrate thereof lie withinthe range of from about 0.1 mg to about 1000 mg per day, given as asingle once-a-day dose in the morning or as divided doses throughout theday taken with food. Particular dosage forms can have about 0.1, 0.2,0.3, 0.4, 0.5, 1.0, 2.0, 2.5, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 100,200, 250, 500, or 1000 mg of the active compound.

Oral Dosage Forms

Pharmaceutical compositions that are suitable for oral administrationcan be presented as discrete dosage forms, such as, but are not limitedto, tablets (e.g., chewable tablets), caplets, capsules, and liquids(e.g., flavored syrups). Such dosage forms contain predetermined amountsof active ingredients, and may be prepared by methods of pharmacy wellknown to those skilled in the art. See generally, Remington'sPharmaceutical Sciences, 20th ed., Mack Publishing, Easton Pa. (2000).

In certain embodiments, the oral dosage forms are solid and preparedunder anhydrous conditions with anhydrous ingredients, as described indetail herein. However, the scope of the compositions provided hereinextends beyond anhydrous, solid oral dosage forms. As such, furtherforms are described herein.

Typical oral dosage forms are prepared by combining the activeingredient(s) in an intimate admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or non-aqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms include,but are not limited to, binders, fillers, disintegrants, and lubricants.Binders suitable for use in pharmaceutical compositions and dosage formsinclude, but are not limited to, corn starch, potato starch, or otherstarches, gelatin, natural and synthetic gums such as acacia, sodiumalginate, alginic acid, other alginates, powdered tragacanth, guar gum,cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,carboxymethyl cellulose calcium, sodium carboxymethyl cellulose),polyvinyl pyrrolidone, methyl cellulose, pre gelatinized starch,hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910),microcrystalline cellulose, and mixtures thereof.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions is typically presentin from about 50 to about 99 weight percent of the pharmaceuticalcomposition or dosage form.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL PH 101, AVICEL PH 103 AVICEL RC581, AVICEL PH 105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Aspecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC 581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL PH 103™ and Starch 1500LM.

Disintegrants are used in the compositions to provide tablets thatdisintegrate when exposed to an aqueous environment. Tablets thatcontain too much disintegrant may disintegrate in storage, while thosethat contain too little may not disintegrate at a desired rate or underthe desired conditions. Thus, a sufficient amount of disintegrant thatis neither too much nor too little to detrimentally alter the release ofthe active ingredients should be used to form solid oral dosage forms.The amount of disintegrant used varies based upon the type offormulation, and is readily discernible to those of ordinary skill inthe art. Typical pharmaceutical compositions comprise from about 0.5 toabout 15 weight percent of disintegrant, specifically from about 1 toabout 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms include, but are not limited to, agar, alginic acid, calciumcarbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polacrilin potassium, sodium starch glycolate, potato ortapioca starch, pre gelatinized starch, other starches, clays, otheralgins, other celluloses, gums, and mixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms include, but are not limited to, calcium stearate, magnesiumstearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol,polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate,talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil,sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zincstearate, ethyl oleate, ethyl laureate, agar, and mixtures thereof.Additional lubricants include, for example, a syloid silica gel (AEROSIL200, manufactured by W. R. Grace Co. of Baltimore, Md.), a coagulatedaerosol of synthetic silica (marketed by Degussa Co. of Plano, Tex.),CAB O SIL (a pyrogenic silicon dioxide product sold by Cabot Co. ofBoston, Mass.), and mixtures thereof. If used at all, lubricants aretypically used in an amount of less than about 1 weight percent of thepharmaceutical compositions or dosage forms into which they areincorporated.

Delayed Release Dosage Forms

Active ingredients such as the compounds provided herein can beadministered by controlled release means or by delivery devices that arewell known to those of ordinary skill in the art. Examples include, butare not limited to, those described in U.S. Pat. Nos. 3,845,770;3,916,899; 3,536,809; 3,598,123; and 4,008,719; 5,674,533; 5,059,595;5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,639,480;5,733,566; 5,739,108; 5,891,474; 5,922,356; 5,972,891; 5,980,945;5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350; 6,248,363;6,264,970; 6,267,981; 6,376,461; 6,419,961; 6,589,548; 6,613,358; and6,699,500; each of which is incorporated herein by reference in itsentirety. Such dosage forms can be used to provide slow or controlledrelease of one or more active ingredients using, for example,hydropropylmethyl cellulose, other polymer matrices, gels, permeablemembranes, osmotic systems, multilayer coatings, microparticles,liposomes, microspheres, or a combination thereof to provide the desiredrelease profile in varying proportions. Suitable controlled releaseformulations known to those of ordinary skill in the art, includingthose described herein, can be readily selected for use with the activeingredients provided herein. Thus encompassed herein are single unitdosage forms suitable for oral administration such as, but not limitedto, tablets, capsules, gelcaps, and caplets that are adapted forcontrolled release.

All controlled release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designed controlledrelease preparation in medical treatment is characterized by a minimumof drug substance being employed to cure or control the condition in aminimum amount of time. Advantages of controlled release formulationsinclude extended activity of the drug, reduced dosage frequency, andincreased subject compliance. In addition, controlled releaseformulations can be used to affect the time of onset of action or othercharacteristics, such as blood levels of the drug, and can thus affectthe occurrence of side (e.g., adverse) effects.

Most controlled release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

In certain embodiments, the drug may be administered using intravenousinfusion, an implantable osmotic pump, a transdermal patch, liposomes,or other modes of administration. In certain embodiments, a pump may beused (see, Sefton, CRC Crit. Ref Biomed. Eng. 14:201 (1987); Buchwald etal., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574(1989)). In another embodiment, polymeric materials can be used. In yetanother embodiment, a controlled release system can be placed in asubject at an appropriate site determined by a practitioner of skill,i.e., thus requiring only a fraction of the systemic dose (see, e.g.,Goodson, Medical Applications of Controlled Release, vol. 2, pp. 115-138(1984)). Other controlled release systems are discussed in the review byLanger (Science 249:1527-1533 (1990)). The active ingredient can bedispersed in a solid inner matrix, e.g., polymethylmethacrylate,polybutylmethacrylate, plasticized or unplasticized polyvinylchloride,plasticized nylon, plasticized polyethyleneterephthalate, naturalrubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene,ethylene-vinylacetate copolymers, silicone rubbers,polydimethylsiloxanes, silicone carbonate copolymers, hydrophilicpolymers such as hydrogels of esters of acrylic and methacrylic acid,collagen, cross-linked polyvinylalcohol and cross-linked partiallyhydrolyzed polyvinyl acetate, that is surrounded by an outer polymericmembrane, e.g., polyethylene, polypropylene, ethylene/propylenecopolymers, ethylene/ethyl acrylate copolymers, ethylene/vinylacetatecopolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber,chlorinated polyethylene, polyvinylchloride, vinylchloride copolymerswith vinyl acetate, vinylidene chloride, ethylene and propylene, ionomerpolyethylene terephthalate, butyl rubber epichlorohydrin rubbers,ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcoholterpolymer, and ethylene/vinyloxyethanol copolymer, that is insoluble inbody fluids. The active ingredient then diffuses through the outerpolymeric membrane in a release rate controlling step. The percentage ofactive ingredient in such parenteral compositions is highly dependent onthe specific nature thereof, as well as the needs of the subject.

Parenteral Dosage Forms

In certain embodiments, provided are parenteral dosage forms. Parenteraldosage forms can be administered to subjects by various routesincluding, but not limited to, subcutaneous, intravenous (includingbolus injection), intramuscular, and intraarterial. Because theiradministration typically bypasses subjects' natural defenses againstcontaminants, parenteral dosage forms are typically, sterile or capableof being sterilized prior to administration to a subject. Examples ofparenteral dosage forms include, but are not limited to, solutions readyfor injection, dry products ready to be dissolved or suspended in apharmaceutically acceptable vehicle for injection, suspensions ready forinjection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage formsare well known to those skilled in the art. Examples include, but arenot limited to: Water for Injection USP; aqueous vehicles such as, butnot limited to, Sodium Chloride Injection, Ringer's Injection, DextroseInjection, Dextrose and Sodium Chloride Injection, and Lactated Ringer'sInjection; water miscible vehicles such as, but not limited to, ethylalcohol, polyethylene glycol, and polypropylene glycol; and non-aqueousvehicles such as, but not limited to, corn oil, cottonseed oil, peanutoil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms.

Transdermal, Topical & Mucosal Dosage Forms

Also provided are transdermal, topical, and mucosal dosage forms.Transdermal, topical, and mucosal dosage forms include, but are notlimited to, ophthalmic solutions, sprays, aerosols, creams, lotions,ointments, gels, solutions, emulsions, suspensions, or other forms knownto one of skill in the art. See, e.g., Remington's PharmaceuticalSciences, 16^(th), 18th and 20^(th) eds., Mack Publishing, Easton Pa.(1980, 1990 & 2000); and Introduction to Pharmaceutical Dosage Forms,4th ed., Lea & Febiger, Philadelphia (1985). Dosage forms suitable fortreating mucosal tissues within the oral cavity can be formulated asmouthwashes or as oral gels. Further, transdermal dosage forms include“reservoir type” or “matrix type” patches, which can be applied to theskin and worn for a specific period of time to permit the penetration ofa desired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed herein are well known to those skilled in thepharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol, butane 1,3diol, isopropyl myristate, isopropyl palmitate, mineral oil, andmixtures thereof to form lotions, tinctures, creams, emulsions, gels orointments, which are nontoxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences, 16^(th), 18th and 20^(th) eds., MackPublishing, Easton Pa. (1980, 1990 & 2000).

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients provided. For example, penetration enhancers canbe used to assist in delivering the active ingredients to the tissue.Suitable penetration enhancers include, but are not limited to: acetone;various alcohols such as ethanol, oleyl, and tetrahydrofuryl; alkylsulfoxides such as dimethyl sulfoxide; dimethyl acetamide; dimethylformamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery enhancing orpenetration enhancing agent. Different salts, hydrates, or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

Dosage and Unit Dosage Forms

In human therapeutics, the doctor will determine the posology which heconsiders most appropriate according to a preventive or curativetreatment and according to the age, weight, stage of the infection andother factors specific to the subject to be treated. In certainembodiments, doses are from about 1 to about 1000 mg per day for anadult, or from about 5 to about 250 mg per day or from about 10 to 50 mgper day for an adult. In certain embodiments, doses are from about 5 toabout 400 mg per day or 25 to 200 mg per day per adult. In certainembodiments, dose rates of from about 50 to about 500 mg per day arealso contemplated.

In further aspects, provided are methods of treating or preventing anHCV infection in a subject by administering, to a subject in needthereof, an effective amount of a compound provided herein, or apharmaceutically acceptable salt thereof. The amount of the compound orcomposition which will be effective in the prevention or treatment of adisorder or one or more symptoms thereof will vary with the nature andseverity of the disease or condition, and the route by which the activeingredient is administered. The frequency and dosage will also varyaccording to factors specific for each subject depending on the specifictherapy (e.g., therapeutic or prophylactic agents) administered, theseverity of the disorder, disease, or condition, the route ofadministration, as well as age, body, weight, response, and the pastmedical history of the subject. Effective doses may be extrapolated fromdose-response curves derived from in vitro or animal model test systems.

In certain embodiments, exemplary doses of a composition includemilligram or microgram amounts of the active compound per kilogram ofsubject or sample weight (e.g., about 10 micrograms per kilogram toabout 50 milligrams per kilogram, about 100 micrograms per kilogram toabout 25 milligrams per kilogram, or about 100 microgram per kilogram toabout 10 milligrams per kilogram). For compositions provided herein, incertain embodiments, the dosage administered to a subject is 0.140 mg/kgto 3 mg/kg of the subject's body weight, based on weight of the activecompound. In certain embodiments, the dosage administered to a subjectis between 0.20 mg/kg and 2.00 mg/kg, or between 0.30 mg/kg and 1.50mg/kg of the subject's body weight.

In certain embodiments, the recommended daily dose range of acomposition provided herein for the conditions described herein liewithin the range of from about 0.1 mg to about 1000 mg per day, given asa single once-a-day dose or as divided doses throughout a day. Incertain embodiments, the daily dose is administered twice daily inequally divided doses. In certain embodiments, a daily dose range shouldbe from about 10 mg to about 200 mg per day, in other embodiments,between about 10 mg and about 150 mg per day, in further embodiments,between about 25 and about 100 mg per day. It may be necessary to usedosages of the active ingredient outside the ranges disclosed herein insome cases, as will be apparent to those of ordinary skill in the art.Furthermore, it is noted that the clinician or treating physician willknow how and when to interrupt, adjust, or terminate therapy inconjunction with subject response.

Different therapeutically effective amounts may be applicable fordifferent diseases and conditions, as will be readily known by those ofordinary skill in the art. Similarly, amounts sufficient to prevent,manage, treat or ameliorate such disorders, but insufficient to cause,or sufficient to reduce, adverse effects associated with the compositionprovided herein are also encompassed by the herein described dosageamounts and dose frequency schedules. Further, when a subject isadministered multiple dosages of a composition provided herein, not allof the dosages need be the same. For example, the dosage administered tothe subject may be increased to improve the prophylactic or therapeuticeffect of the composition or it may be decreased to reduce one or moreside effects that a particular subject is experiencing.

In certain embodiment, the dosage of the composition provided herein,based on weight of the active compound, administered to prevent, treat,manage, or ameliorate a disorder, or one or more symptoms thereof in asubject is 0.1 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6mg/kg, 10 mg/kg, or 15 mg/kg or more of a subject's body weight. Inanother embodiment, the dosage of the composition or a compositionprovided herein administered to prevent, treat, manage, or ameliorate adisorder, or one or more symptoms thereof in a subject is a unit dose of0.1 mg to 200 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg,0.1 mg to 20 mg, 0.1 mg to 15 mg, 0.1 mg to 10 mg, 0.1 mg to 7.5 mg, 0.1mg to 5 mg, 0.1 to 2.5 mg, 0.25 mg to 20 mg, 0.25 to 15 mg, 0.25 to 12mg, 0.25 to 10 mg, 0.25 mg to 7.5 mg, 0.25 mg to 5 mg, 0.5 mg to 2.5 mg,1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 12 mg, 1 mg to 10 mg, 1 mg to 7.5mg, 1 mg to 5 mg, or 1 mg to 2.5 mg.

In certain embodiments, treatment or prevention can be initiated withone or more loading doses of a compound or composition provided hereinfollowed by one or more maintenance doses. In such embodiments, theloading dose can be, for instance, about 60 to about 400 mg per day, orabout 100 to about 200 mg per day for one day to five weeks. The loadingdose can be followed by one or more maintenance doses. In certainembodiments, each maintenance does is, independently, about from about10 mg to about 200 mg per day, between about 25 mg and about 150 mg perday, or between about 25 and about 80 mg per day. Maintenance doses canbe administered daily and can be administered as single doses, or asdivided doses.

In certain embodiments, a dose of a compound or composition providedherein can be administered to achieve a steady-state concentration ofthe active ingredient in blood or serum of the subject. The steady-stateconcentration can be determined by measurement according to techniquesavailable to those of skill or can be based on the physicalcharacteristics of the subject such as height, weight, and age. Incertain embodiments, a sufficient amount of a compound or compositionprovided herein is administered to achieve a steady-state concentrationin blood or serum of the subject of from about 300 to about 4000 ng/mL,from about 400 to about 1600 ng/mL, or from about 600 to about 1200ng/mL. In some embodiments, loading doses can be administered to achievesteady-state blood or serum concentrations of about 1200 to about 8000ng/mL, or about 2000 to about 4000 ng/mL for one to five days. Incertain embodiments, maintenance doses can be administered to achieve asteady-state concentration in blood or serum of the subject of fromabout 300 to about 4000 ng/mL, from about 400 to about 1600 ng/mL, orfrom about 600 to about 1200 ng/mL.

In certain embodiments, administration of the same composition may berepeated and the administrations may be separated by at least 1 day, 2days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75days, 3 months, or 6 months. In other embodiments, administration of thesame prophylactic or therapeutic agent may be repeated and theadministration may be separated by at least at least 1 day, 2 days, 3days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3months, or 6 months.

In certain aspects, provided herein are unit dosages comprising acompound, or a pharmaceutically acceptable salt thereof, in a formsuitable for administration. Such forms are described in detail herein.In certain embodiments, the unit dosage comprises 1 to 1000 mg, 5 to 250mg, or 10 to 50 mg active ingredient. In particular embodiments, theunit dosages comprise about 1, 5, 10, 25, 50, 100, 125, 250, 500, or1000 mg active ingredient. Such unit dosages can be prepared accordingto techniques familiar to those of skill in the art.

The dosages of the second agents are to be used in the combinationtherapies provided herein. In certain embodiments, dosages lower thanthose which have been or are currently being used to prevent or treatHCV infection are used in the combination therapies provided herein. Therecommended dosages of second agents can be obtained from the knowledgeof those of skill. For those second agents that are approved forclinical use, recommended dosages are described in, for example, Hardmanet al., eds., 1996, Goodman & Gilman's The Pharmacological Basis OfBasis Of Therapeutics 9^(th) Ed, Mc-Graw-Hill, New York; Physician'sDesk Reference (PDR) 57^(th) Ed., 2003, Medical Economics Co., Inc.,Montvale, N.J., which are incorporated herein by reference in itsentirety.

In various embodiments, the therapies (e.g., a compound provided hereinand the second agent) are administered less than 5 minutes apart, lessthan 30 minutes apart, 1 hour apart, at about 1 hour apart, at about 1to about 2 hours apart, at about 2 hours to about 3 hours apart, atabout 3 hours to about 4 hours apart, at about 4 hours to about 5 hoursapart, at about 5 hours to about 6 hours apart, at about 6 hours toabout 7 hours apart, at about 7 hours to about 8 hours apart, at about 8hours to about 9 hours apart, at about 9 hours to about 10 hours apart,at about 10 hours to about 11 hours apart, at about 11 hours to about 12hours apart, at about 12 hours to 18 hours apart, 18 hours to 24 hoursapart, 24 hours to 36 hours apart, 36 hours to 48 hours apart, 48 hoursto 52 hours apart, 52 hours to 60 hours apart, 60 hours to 72 hoursapart, 72 hours to 84 hours apart, 84 hours to 96 hours apart, or 96hours to 120 hours apart. In various embodiments, the therapies areadministered no more than 24 hours apart or no more than 48 hours apart.In certain embodiments, two or more therapies are administered withinthe same patient visit. In other embodiments, the compound providedherein and the second agent are administered concurrently.

In other embodiments, the compound provided herein and the second agentare administered at about 2 to 4 days apart, at about 4 to 6 days apart,at about 1 week part, at about 1 to 2 weeks apart, or more than 2 weeksapart.

In certain embodiments, administration of the same agent may be repeatedand the administrations may be separated by at least 1 day, 2 days, 3days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3months, or 6 months. In other embodiments, administration of the sameagent may be repeated and the administration may be separated by atleast at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days,45 days, 2 months, 75 days, 3 months, or 6 months.

In certain embodiments, a compound provided herein and a second agentare administered to a patient, for example, a mammal, such as a human,in a sequence and within a time interval such that the compound providedherein can act together with the other agent to provide an increasedbenefit than if they were administered otherwise. For example, thesecond active agent can be administered at the same time or sequentiallyin any order at different points in time; however, if not administeredat the same time, they should be administered sufficiently close in timeso as to provide the desired therapeutic or prophylactic effect. Incertain embodiments, the compound provided herein and the second activeagent exert their effect at times which overlap. Each second activeagent can be administered separately, in any appropriate form and by anysuitable route. In other embodiments, the compound provided herein isadministered before, concurrently or after administration of the secondactive agent.

In certain embodiments, the compound provided herein and the secondagent are cyclically administered to a patient. Cycling therapy involvesthe administration of a first agent (e.g., a first prophylactic ortherapeutic agents) for a period of time, followed by the administrationof a second agent and/or third agent (e.g., a second and/or thirdprophylactic or therapeutic agents) for a period of time and repeatingthis sequential administration. Cycling therapy can reduce thedevelopment of resistance to one or more of the therapies, avoid orreduce the side effects of one of the therapies, and/or improve theefficacy of the treatment.

In certain embodiments, the compound provided herein and the secondactive agent are administered in a cycle of less than about 3 weeks,about once every two weeks, about once every 10 days or about once everyweek. One cycle can comprise the administration of a compound providedherein and the second agent by infusion over about 90 minutes everycycle, about 1 hour every cycle, about 45 minutes every cycle. Eachcycle can comprise at least 1 week of rest, at least 2 weeks of rest, atleast 3 weeks of rest. The number of cycles administered is from about 1to about 12 cycles, more typically from about 2 to about 10 cycles, andmore typically from about 2 to about 8 cycles.

In other embodiments, courses of treatment are administered concurrentlyto a patient, i.e., individual doses of the second agent areadministered separately yet within a time interval such that thecompound provided herein can work together with the second active agent.For example, one component can be administered once per week incombination with the other components that can be administered onceevery two weeks or once every three weeks. In other words, the dosingregimens are carried out concurrently even if the therapeutics are notadministered simultaneously or during the same day.

The second agent can act additively or synergistically with the compoundprovided herein. In certain embodiments, the compound provided herein isadministered concurrently with one or more second agents in the samepharmaceutical composition. In another embodiment, a compound providedherein is administered concurrently with one or more second agents inseparate pharmaceutical compositions. In still another embodiment, acompound provided herein is administered prior to or subsequent toadministration of a second agent. Also contemplated are administrationof a compound provided herein and a second agent by the same ordifferent routes of administration, e.g., oral and parenteral. Incertain embodiments, when the compound provided herein is administeredconcurrently with a second agent that potentially produces adverse sideeffects including, but not limited to, toxicity, the second active agentcan advantageously be administered at a dose that falls below thethreshold that the adverse side effect is elicited.

Kits

Also provided are kits for use in methods of treatment of a liverdisorder such as HCV infections. The kits can include a compound orcomposition provided herein, a second agent or composition, andinstructions providing information to a health care provider regardingusage for treating the disorder. Instructions may be provided in printedform or in the form of an electronic medium such as a floppy disc, CD,or DVD, or in the form of a website address where such instructions maybe obtained. A unit dose of a compound or composition provided herein,or a second agent or composition, can include a dosage such that whenadministered to a subject, a therapeutically or prophylacticallyeffective plasma level of the compound or composition can be maintainedin the subject for at least 1 days. In some embodiments, a compound orcomposition can be included as a sterile aqueous pharmaceuticalcomposition or dry powder (e.g., lyophilized) composition.

In some embodiments, suitable packaging is provided. As used herein,“packaging” includes a solid matrix or material customarily used in asystem and capable of holding within fixed limits a compound providedherein and/or a second agent suitable for administration to a subject.Such materials include glass and plastic (e.g., polyethylene,polypropylene, and polycarbonate) bottles, vials, paper, plastic, andplastic-foil laminated envelopes and the like. If e-beam sterilizationtechniques are employed, the packaging should have sufficiently lowdensity to permit sterilization of the contents.

Methods of Use

Provided herein is a method for inhibiting replication of a virus in ahost, which comprises contacting the host with a therapeuticallyeffective amount of a thiophosphate nucleoside disclosed herein, e.g., athiophosphate nucleoside compound of Formula 2001, I-XXIX, 1-23byii and1001-1002, including a single enantiomer, a mixture of an enantiomericpair, an individual diastereomer, a mixture of diastereomers, or atautomeric form thereof or a pharmaceutically acceptable salt, solvate,prodrug, phosphate, or active metabolite thereof.

Provided herein is a method for inhibiting replication of a virus in acell, which comprises contacting the cell with a therapeuticallyeffective amount of a thiophosphate nucleoside compound disclosedherein, e.g., a thiophosphate nucleoside compound of Formula 2001,I-XXIX, 1-23byii and 1001-1002, including a single enantiomer, a mixtureof an enantiomeric pair, an individual diastereomer, a mixture ofdiastereomers, or a tautomeric form thereof; or a pharmaceuticallyacceptable salt, solvate, prodrug, phosphate, or active metabolitethereof.

Provided herein is a method for inhibiting replication of a virus, whichcomprises contacting the virus with a therapeutically effective amountof a thiophosphate nucleoside compound disclosed herein, e.g., athiophosphate nucleoside compound of Formula 2001, I-XXIX, 1-23byii and1001-1002, including a single enantiomer, a mixture of an enantiomericpair, an individual diastereomer, a mixture of diastereomers, or atautomeric form thereof; or a pharmaceutically acceptable salt, solvate,prodrug, phosphate, or active metabolite thereof.

Provided herein is a method for inhibiting the activity of a polymerase,which comprises contacting the polymerase with a ester or malonate of athiophosphate nucleoside compound disclosed herein, e.g., athiophosphate nucleoside compound of Formula 2001, I-XXIX, 1-23byii and1001-1002, including a single enantiomer, a mixture of an enantiomericpair, an individual diastereomer, a mixture of diastereomers, or atautomeric form thereof; or a pharmaceutically acceptable salt, solvate,prodrug, phosphate, or active metabolite thereof.

In certain embodiments, provided herein are methods for the treatmentand/or prophylaxis of a host infected with Flaviviridae that includesthe administration of an effective amount of a thiophosphate nucleosidecompound disclosed herein, e.g., a thiophosphate nucleoside compound ofFormula 2001, I-XXIX, 1-23byii and 1001-1002, including a singleenantiomer, a mixture of an enantiomeric pair, an individualdiastereomer, a mixture of diastereomers, or a tautomeric form thereof;or a pharmaceutically acceptable salt, solvate, prodrug, phosphate, oractive metabolite thereof.

In certain embodiments, provided herein are methods for treating an HCVinfection in a subject. In certain embodiments, the methods encompassthe step of administering to a subject in need thereof an amount of acompound effective for the treatment or prevention of an HCV infectionin combination with a second agent effective for the treatment orprevention of the infection. The compound can be any compound asdescribed herein, and the second agent can be any second agent describedin the art or herein. In certain embodiments, the compound is in theform of a pharmaceutical composition or dosage form, as describedelsewhere herein.

In certain embodiments, provided herein are methods for the treatmentand/or prophylaxis of a host infected with Flaviviridae that includesthe administration of an effective amount of a compounds providedherein, or a pharmaceutically acceptable salt thereof. In certainembodiments, provided herein are methods for treating an HCV infectionin a subject. In certain embodiments, the methods encompass the step ofadministering to a subject in need thereof an amount of a compoundeffective for the treatment or prevention of an HCV infection incombination with a second agent effective for the treatment orprevention of the infection. The compound can be any compound asdescribed herein, and the second agent can be any second agent describedin the art or herein. In certain embodiments, the compound is in theform of a pharmaceutical composition or dosage form, as describedelsewhere herein.

Flaviviridae which can be treated are, e.g., discussed generally inFields Virology, Fifth Ed., Editors: Knipe, D. M., and Howley, P. M.,Lippincott Williams & Wilkins Publishers, Philadelphia, Pa., Chapters33-35, 2006. In a particular embodiment of the invention, theFlaviviridae is HCV. In an alternate embodiment, the Flaviviridae is aflavivirus or pestivirus. In certain embodiments, the Flaviviridae canbe from any class of Flaviviridae. In certain embodiments, theFlaviviridae is a mammalian tick-borne virus. In certain embodiments,the Flaviviridae is a seabird tick-borne virus. In certain embodiments,the Flaviviridae is a mosquito-borne virus. In certain embodiments, theFlaviviridae is an Aroa virus. In certain embodiments, the Flaviviridaeis a Dengue virus. In certain embodiments, the Flaviviridae is aJapanese encephalitis virus. In certain embodiments, the Flaviviridae isa Kokobera virus. In certain embodiments, the Flaviviridae is a Ntayavirus. In certain embodiments, the Flaviviridae is a Spondweni virus. Incertain embodiments, the Flaviviridae is a Yellow fever virus. Incertain embodiments, the Flaviviridae is a Entebbe virus. In certainembodiments, the Flaviviridae is a Modoc virus. In certain embodiments,the Flaviviridae is a Rio Bravo virus.

Specific flaviviruses which can be treated include, without limitation:Absettarov, Aedes, Alfuy, Alkhurma, Apoi, Aroa, Bagaza, Banzi, Bukalasabat, Bouboui, Bussuquara, Cacipacore, Calbertado, Carey Island, Cellfusing agent, Cowbone Ridge, Culex, Dakar bat, Dengue 1, Dengue 2,Dengue 3, Dengue 4, Edge Hill, Entebbe bat, Gadgets Gully, Hanzalova,Hypr, Ilheus, Israel turkey meningoencephalitis, Japanese encephalitis,Jugra, Jutiapa, Kadam, Kamiti River, Karshi, Kedougou, Kokobera,Koutango, Kumlinge, Kunjin, Kyasanur Forest disease, Langat, Loupingill, Meaban, Modoc, Montana myotis leukoencephalitis, Murray valleyencephalitis, Nakiwogo, Naranjal, Negishi, Ntaya, Omsk hemorrhagicfever, Phnom-Penh bat, Powassan, Quang Binh, Rio Bravo, Rocio, RoyalFarm, Russian spring-summer encephalitis, Saboya, St. Louisencephalitis, Sal Vieja, San Perlita, Saumarez Reef, Sepik, Sokuluk,Spondweni, Stratford, Tembusu, Tick-borne encephalitis, Turkish sheepencephalitis, Tyuleniy, Uganda S, Usutu, Wesselsbron, West Nile,Yaounde, Yellow fever, Yokose, and Zika.

Pestiviruses which can be treated are discussed generally in FieldsVirology, Fifth Ed., Editors: Knipe, D. M., and Howley, P. M.,Lippincott Williams & Wilkins Publishers, Philadelphia, Pa., Chapters33-35, 2006. Specific pestiviruses which can be treated include, withoutlimitation: bovine viral diarrhea virus (“BVDV”), classical swine fevervirus (“CSFV,” also called hog cholera virus), and border disease virus(“BDV”).

In certain embodiments, the subject can be any subject infected with, orat risk for infection with, HCV. Infection or risk for infection can bedetermined according to any technique deemed suitable by thepractitioner of skill in the art. In certain embodiments, subjects arehumans infected with HCV.

In certain embodiments, the subject has never received therapy orprophylaxis for an HCV infection. In further embodiments, the subjecthas previously received therapy or prophylaxis for an HCV infection. Forinstance, in certain embodiments, the subject has not responded to anHCV therapy. For example, under current interferon therapy, up to 50% ormore HCV subjects do not respond to therapy. In certain embodiments, thesubject can be a subject that received therapy but continued to sufferfrom viral infection or one or more symptoms thereof. In certainembodiments, the subject can be a subject that received therapy butfailed to achieve a sustained virologic response. In certainembodiments, the subject has received therapy for an HCV infection buthas failed to show, for example, a 2 log₁₀ decline in HCV RNA levelsafter 12 weeks of therapy. It is believed that subjects who have notshown more than 2 log₁₀ reduction in serum HCV RNA after 12 weeks oftherapy have a 97-100% chance of not responding.

In certain embodiments, the subject is a subject that discontinued anHCV therapy because of one or more adverse events associated with thetherapy. In certain embodiments, the subject is a subject where currenttherapy is not indicated. For instance, certain therapies for HCV areassociated with neuropsychiatric events. Interferon (IFN)-alfa plusribavirin is associated with a high rate of depression. Depressivesymptoms have been linked to a worse outcome in a number of medicaldisorders. Life-threatening or fatal neuropsychiatric events, includingsuicide, suicidal and homicidal ideation, depression, relapse of drugaddiction/overdose, and aggressive behavior have occurred in subjectswith and without a previous psychiatric disorder during HCV therapy.Interferon-induced depression is a limitation for the treatment ofchronic hepatitis C, especially for subjects with psychiatric disorders.Psychiatric side effects are common with interferon therapy andresponsible for about 10% to 20% of discontinuations of current therapyfor HCV infection.

Accordingly, provided are methods of treating or preventing an HCVinfection in subjects where the risk of neuropsychiatric events, such asdepression, contraindicates treatment with current HCV therapy. Incertain embodiments, provided are methods of treating or preventing HCVinfection in subjects where a neuropsychiatric event, such asdepression, or risk of such indicates discontinuation of treatment withcurrent HCV therapy. Further provided are methods of treating orpreventing HCV infection in subjects where a neuropsychiatric event,such as depression, or risk of such indicates dose reduction of currentHCV therapy.

Current therapy is also contraindicated in subjects that arehypersensitive to interferon or ribavirin, or both, or any othercomponent of a pharmaceutical product for administration of interferonor ribavirin. Current therapy is not indicated in subjects withhemoglobinopathies (e.g., thalassemia major, sickle-cell anemia) andother subjects at risk from the hematologic side effects of currenttherapy. Common hematologic side effects include bone marrowsuppression, neutropenia, and thrombocytopenia. Furthermore, ribavirinis toxic to red blood cells and is associated with hemolysis.Accordingly, in certain embodiments, provided are methods of treating orpreventing HCV infection in subjects hypersensitive to interferon orribavirin, or both, subjects with a hemoglobinopathy, for instancethalassemia major subjects and sickle-cell anemia subjects, and othersubjects at risk from the hematologic side effects of current therapy.

In certain embodiments, the subject has received an HCV therapy anddiscontinued that therapy prior to administration of a method providedherein. In further embodiments, the subject has received therapy andcontinues to receive that therapy along with administration of a methodprovided herein. The methods can be co-administered with other therapyfor HBC and/or HCV according to the judgment of one of skill in the art.In certain embodiments, the methods or compositions provided herein canbe co-administered with a reduced dose of the other therapy for HBCand/or HCV.

In certain embodiments, provided are methods of treating a subject thatis refractory to treatment with interferon. For instance, in someembodiments, the subject can be a subject that has failed to respond totreatment with one or more agents, wherein the one or more agents are aninterferon, interferon α, pegylated interferon α, interferon plusribavirin, interferon α plus ribavirin, and/or pegylated interferon αplus ribavirin. In some embodiments, the subject can be a subject thathas responded poorly to treatment with one or more agents, wherein theone or more agents are interferon, interferon α, pegylated interferon α,interferon plus ribavirin, interferon α plus ribavirin, and/or pegylatedinterferon α plus ribavirin. A pro-drug form of ribavirin, such astaribavirin, may also be used.

In certain embodiments, the subject has, or is at risk for, co-infectionof HCV with HIV. For instance, in the United States, 30% of HIV subjectsare co-infected with HCV and evidence indicates that people infectedwith HIV have a much more rapid course of their hepatitis C infection.Maier and Wu, 2002, World J Gastroenterol 8:577-57. The methods providedherein can be used to treat or prevent HCV infection in such subjects.It is believed that elimination of HCV in these subjects will lowermortality due to end-stage liver disease. Indeed, the risk ofprogressive liver disease is higher in subjects with severeAIDS-defining immunodeficiency than in those without. See, e.g., Lesenset al., 1999, J Infect Dis 179:1254-1258. In certain embodiments,compounds provided herein have been shown to suppress HIV in HIVsubjects. Thus, in certain embodiments, provided are methods of treatingor preventing HIV infection and HCV infection in subjects in needthereof.

In certain embodiments, the compounds or compositions are administeredto a subject following liver transplant. Hepatitis C is a leading causeof liver transplantation in the U.S., and many subjects that undergoliver transplantation remain HCV positive following transplantation. Incertain embodiments, provided are methods of treating such recurrent HCVsubjects with a compound or composition provided herein. In certainembodiments, provided are methods of treating a subject before, duringor following liver transplant to prevent recurrent HCV infection.

Assay Methods

Compounds can be assayed for HCV activity according to any assay knownto those of skill in the art.

Further, compounds can be assayed for accumulation in liver cells of asubject according to any assay known to those of skill in the art. Incertain embodiments, a compound can be administered to the subject, anda liver cell of the subject can be assayed for the compound or aderivative thereof, e.g. a nucleoside, nucleoside phosphate ornucleoside triphosphate derivative thereof.

In certain embodiments, a thiophosphate nucleoside compound isadministered to cells, such as liver cells, in vivo or in vitro, and thenucleoside triphosphate levels delivered intracellularly are measured,to indicate delivery of the compound and triphosphorylation in the cell.The levels of intracellular nucleoside triphosphate can be measuredusing analytical techniques known in the art. Methods of detecting ddATPare described herein below by way of example, but other nucleosidetriphosphates can be readily detected using the appropriate controls,calibration samples, and assay techniques.

In certain embodiments, ddATP concentrations are measured in a sample bycomparison to calibration standards made from control samples. The ddATPconcentrations in a sample can be measured using an analytical methodsuch as HPLC LC MS. In certain embodiments, a test sample is compared toa calibration curve created with known concentrations of ddATP tothereby obtain the concentration of that sample.

In certain embodiments, the samples are manipulated to remove impuritiessuch as salts (Na⁺, K⁺, etc.) before analysis. In certain embodiments,the lower limit of quantitation is about ˜0.2 pmol/mL for hepatocytecellular extracts particularly where reduced salt is present.

In certain embodiments, the method allows successfully measuringtriphosphate nucleotides formed at levels of 1-10,000 pmol per millioncells in e.g. cultured hepatocytes and HepG2 cells.

Second Therapeutic Agents

In certain embodiments, the compounds and compositions provided hereinare useful in methods of treatment of a liver disorder, that comprisefurther administration of a second agent effective for the treatment ofthe disorder, such as HCV infection in a subject in need thereof. Thesecond agent can be any agent known to those of skill in the art to beeffective for the treatment of the disorder, including those currentlyapproved by the FDA.

In certain embodiments, a compound provided herein is administered incombination with one second agent. In further embodiments, a secondagent is administered in combination with two second agents. In stillfurther embodiments, a second agent is administered in combination withtwo or more second agents.

As used herein, the term “in combination” includes the use of more thanone therapy (e.g., one or more prophylactic and/or therapeutic agents).The use of the term “in combination” does not restrict the order inwhich therapies (e.g., prophylactic and/or therapeutic agents) areadministered to a subject with a disorder. A first therapy (e.g., aprophylactic or therapeutic agent such as a compound provided herein)can be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes,45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6weeks, 8 weeks, or 12 weeks before), concomitantly with, or subsequentto (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or12 weeks after) the administration of a second therapy (e.g., aprophylactic or therapeutic agent) to a subject with a disorder.

As used herein, the term “synergistic” includes a combination of acompound provided herein and another therapy (e.g., a prophylactic ortherapeutic agent) which has been or is currently being used to prevent,manage, or treat a disorder, which is more effective than the additiveeffects of the therapies. A synergistic effect of a combination oftherapies (e.g., a combination of prophylactic or therapeutic agents)permits the use of lower dosages of one or more of the therapies and/orless frequent administration of said therapies to a subject with adisorder. The ability to utilize lower dosages of a therapy (e.g., aprophylactic or therapeutic agent) and/or to administer said therapyless frequently reduces the toxicity associated with the administrationof said therapy to a subject without reducing the efficacy of saidtherapy in the prevention or treatment of a disorder). In addition, asynergistic effect can result in improved efficacy of agents in theprevention or treatment of a disorder. Finally, a synergistic effect ofa combination of therapies (e.g., a combination of prophylactic ortherapeutic agents) may avoid or reduce adverse or unwanted side effectsassociated with the use of either therapy alone.

The active compounds provided herein can be administered in combinationor alternation with another therapeutic agent, in particular an anti-HCVagent. In combination therapy, effective dosages of two or more agentsare administered together, whereas in alternation or sequential-steptherapy, an effective dosage of each agent is administered serially orsequentially. The dosages given will depend on absorption, inactivation,and excretion rates of the drug as well as other factors known to thoseof skill in the art. It is to be noted that dosage values will also varywith the severity of the condition to be alleviated. It is to be furtherunderstood that for any particular subject, specific dosage regimens andschedules should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the compositions. In certain embodiments, ananti-HCV (or anti-pestivirus or anti-flavivirus) compound that exhibitsan EC₅₀ of 10-15 μM. In certain embodiments, less than 1-5 μM, isdesirable.

It has been recognized that drug-resistant variants of flaviviruses,pestiviruses, or HCV can emerge after prolonged treatment with anantiviral agent. Drug resistance most typically occurs by mutation of agene that encodes for an enzyme used in viral replication. The efficacyof a drug against the viral infection can be prolonged, augmented, orrestored by administering the compound in combination or alternationwith a second, and perhaps third, antiviral compound that induces adifferent mutation from that caused by the principle drug.Alternatively, the pharmacokinetics, biodistribution or other parameterof the drug can be altered by such combination or alternation therapy.In general, combination therapy is typically preferred over alternationtherapy because it induces multiple simultaneous stresses on the virus.

Any of the viral treatments described in the Background of the Inventioncan be used in combination or alternation with the compounds describedin this specification. Non-limiting examples of second agents include:

HCV Protease inhibitors: Examples include Medivir HCV Protease Inhibitorsimeprevir (HCV-PI or TMC435) (Medivir/Tibotec); MK-7009 (Merck), RG7227(ITMN-191) (Roche/Pharmasset/InterMune), boceprevir (SCH 503034)(Schering), SCH 446211 (Schering), narlaprevir SCH900518(Schering/Merck), ABT-450 (Abbott/Enanta), ACH-1625 (Achillion), BI201335 (Boehringer Ingelheim), PHX1766 (Phenomix), VX-500 (Vertex) andtelaprevir (VX-950) (Vertex). Further examples of protease inhibitorsinclude substrate-based NS3 protease inhibitors (Attwood et al.,Antiviral peptide derivatives, PCT WO 98/22496, 1998; Attwood et al.,Antiviral Chemistry and Chemotherapy 1999, 10, 259-273; Attwood et al.,Preparation and use of amino acid derivatives as anti-viral agents,German Patent Pub. DE 19914474; Tung et al., Inhibitors of serineproteases, particularly hepatitis C virus NS3 protease, PCT WO98/17679), including alphaketoamides and hydrazinoureas, and inhibitorsthat terminate in an electrophile such as a boronic acid or phosphonate(Llinas-Brunet et al, Hepatitis C inhibitor peptide analogues, PCT WO99/07734); Non-substrate-based NS3 protease inhibitors such as2,4,6-trihydroxy-3-nitro-benzamide derivatives (Sudo K. et al.,Biochemical and Biophysical Research Communications, 1997, 238, 643-647;Sudo K. et al., Antiviral Chemistry and Chemotherapy, 1998, 9, 186),including RD3-4082 and RD3-4078, the former substituted on the amidewith a 14 carbon chain and the latter processing a para-phenoxyphenylgroup; and Sch 68631, a phenanthrenequinone, an HCV protease inhibitor(Chu M. et al., Tetrahedron Letters 37:7229-7232, 1996).

SCH 351633, isolated from the fungus Penicillium griseofulvum, wasidentified as a protease inhibitor (Chu M. et al., Bioorganic andMedicinal Chemistry Letters 9:1949-1952). Eglin c, isolated from leech,is a potent inhibitor of several serine proteases such as S. griseusproteases A and B, α-chymotrypsin, chymase and subtilisin. Qasim M. A.et al., Biochemistry 36:1598-1607, 1997.

U.S. patents disclosing protease inhibitors for the treatment of HCVinclude, for example, U.S. Pat. No. 6,004,933 to Spruce et al., whichdiscloses a class of cysteine protease inhibitors for inhibiting HCVendopeptidase 2; U.S. Pat. No. 5,990,276 to Zhang et al., whichdiscloses synthetic inhibitors of hepatitis C virus NS3 protease; U.S.Pat. No. 5,538,865 to Reyes et a; WO 02/008251 to Corvas International,Inc., and U.S. Pat. No. 7,169,760, US2005/176648, WO 02/08187 and WO02/008256 to Schering Corporation. HCV inhibitor tripeptides aredisclosed in U.S. Pat. Nos. 6,534,523, 6,410,531, and U.S. Pat. No.6,420,380 to Boehringer Ingelheim and WO 02/060926 to Bristol MyersSquibb. Diaryl peptides as NS3 serine protease inhibitors of HCV aredisclosed in WO 02/48172 and U.S. Pat. No. 6,911,428 to ScheringCorporation. Imidazoleidinones as NS3 serine protease inhibitors of HCVare disclosed in WO 02/08198 and U.S. Pat. No. 6,838,475 to ScheringCorporation and WO 02/48157 and U.S. Pat. No. 6,727,366 to Bristol MyersSquibb. WO 98/17679 and U.S. Pat. No. 6,265,380 to VertexPharmaceuticals and WO 02/48116 and U.S. Pat. No. 6,653,295 to BristolMyers Squibb also disclose HCV protease inhibitors. Further examples ofHCV serine protease inhibitors are provided in U.S. Pat. No. 6,872,805(Bristol-Myers Squibb); WO 2006000085 (Boehringer Ingelheim); U.S. Pat.No. 7,208,600 (Vertex); US 2006/0046956 (Schering-Plough); WO2007/001406 (Chiron); US 2005/0153877; WO 2006/119061 (Merck); WO00/09543 (Boehringer Ingelheim), U.S. Pat. No. 6,323,180 (BoehringerIngelheim) WO 03/064456 (Boehringer Ingelheim), U.S. Pat. No. 6,642,204(Boehringer Ingelheim), WO 03/064416 (Boehringer Ingelheim), U.S. Pat.No. 7,091,184 (Boehringer Ingelheim), WO 03/053349 (Bristol-MyersSquibb), U.S. Pat. No. 6,867,185, WO 03/099316 (Bristol-Myers Squibb),U.S. Pat. No. 6,869,964, WO 03/099274 (Bristol-Myers Squibb), U.S. Pat.No. 6,995,174, WO 2004/032827 (Bristol-Myers Squibb), U.S. Pat. No.7,041,698, WO 2004/043339 and U.S. Pat. No. 6,878,722 (Bristol-MyersSquibb).

Thiazolidine derivatives which show relevant inhibition in areverse-phase HPLC assay with an NS3/4A fusion protein and NS5A/5Bsubstrate (Sudo K. et al., Antiviral Research, 1996, 32, 9-18),especially compound RD-1-6250, possessing a fused cinnamoyl moietysubstituted with a long alkyl chain, RD4 6205 and RD4 6193;

Thiazolidines and benzanilides identified in Kakiuchi N. et al., J. EBSLetters 421, 217-220; Takeshita N. et al., Analytical Biochemistry,1997, 247, 242-246;

A phenanthrenequinone possessing activity against protease in a SDS-PAGEand autoradiography assay isolated from the fermentation culture brothof Streptomyces sp., SCH 68631 (Chu M. et al., Tetrahedron Letters,1996, 37, 7229-7232), and SCH 351633, isolated from the fungusPenicillium griseofulvum, which demonstrates activity in a scintillationproximity assay (Chu M. et al., Bioorganic and Medicinal ChemistryLetters 9, 1949-1952);

Helicase inhibitors (Diana G. D. et al., Compounds, compositions andmethods for treatment of hepatitis C, U.S. Pat. No. 5,633,358; Diana G.D. et al., Piperidine derivatives, pharmaceutical compositions thereofand their use in the treatment of hepatitis C, PCT WO 97/36554);

HCV polymerase inhibitors, including nucleoside and non-nucleosidepolymerase inhibitors, such as ribavirin, viramidine, clemizole,filibuvir (PF-00868554), HCV POL, NM 283 (valopicitabine), MK-0608,7-Fluoro-MK-0608, MK-3281, IDX-375, ABT-072, ABT-333, 0 598, BI 207127,GS 9190, PSI-6130, R1626, PSI-6206, PSI-938, PSI-7851, PSI-7977(sofosbuvir, Sovaldi), RG1479, RG7128, HCV-796 VCH-759 or VCH-916;

Gliotoxin (Ferrari R. et al., Journal of Virology, 1999, 73, 1649-1654),and the natural product cerulenin (Lohmann V. et al., Virology, 1998,249, 108-118);

Interfering RNA (iRNA) based antivirals, including short interfering RNA(siRNA) based antivirals, such as Sirna-034 and others described inInternational Patent Publication Nos. WO/03/070750 and WO 2005/012525,and US Patent Publication No. US 2004/0209831;

HCV NS5A inhibitors, such as BMS-790052 (daclatasvir, Bristol-MyersSquibb), PPI-461 (Presidio Pharmaceuticals), PPI-1301 (PresidioPharmaceuticals), IDX-719 (samatasvir, Idenix Pharmaceuticals), AZD7295(Arrow Therapeutics, AstraZeneca), EDP-239 (Enanta), ACH-2928(Achillion), ACH-3102 (Achillion), ABT-267 (Abbott), or GS-5885(Gilead);

Antisense phosphorothioate oligodeoxynucleotides (S-ODN) complementaryto sequence stretches in the 5′ non-coding region (NCR) of the virus(Alt M. et al., Hepatology, 1995, 22, 707-717), or nucleotides 326-348comprising the 3′ end of the NCR and nucleotides 371-388 located in thecore coding region of the HCV RNA (Alt M. et al., Archives of Virology,1997, 142, 589-599; Galderisi U. et al., Journal of Cellular Physiology,1999, 181, 251-257);

Inhibitors of IRES-dependent translation (Ikeda N et al., Agent for theprevention and treatment of hepatitis C, Japanese Patent Pub.JP-08268890; Kai Y. et al., Prevention and treatment of viral diseases,Japanese Patent Pub. JP-10101591);

HCV entry inhibitors, such as celgosivir (MK-3253) (MIGENIX Inc.), SP-30(Samaritan Pharmaceuticals), ITX4520 (iTherX), ITX5061 (iTherX), PRO-206(Progenics Pharmaceuticals) and other entry inhibitors by ProgenicsPharmaceuticals, e.g., as disclosed in U.S. Patent Publication No.2006/0198855.

Ribozymes, such as nuclease-resistant ribozymes (Maccjak, D. J. et al.,Hepatology 1999, 30, abstract 995) and those disclosed in U.S. Pat. No.6,043,077 to Barber et al., and U.S. Pat. Nos. 5,869,253 and 5,610,054to Draper et al.; and

Nucleoside analogs have also been developed for the treatment ofFlaviviridae infections.

In certain embodiments, the compounds provided herein can beadministered in combination with any of the compounds described byIdenix Pharmaceuticals in International Publication Nos. WO 01/90121, WO01/92282, WO 2004/003000, WO 2004/002422 and WO 2004/002999.

Other patent applications disclosing the use of certain nucleosideanalogs that can be used as second agents to treat hepatitis C virusinclude: PCT/CA00/01316 (WO 01/32153; filed Nov. 3, 2000) andPCT/CA01/00197 (WO 01/60315; filed Feb. 19, 2001) filed by BioChemPharma, Inc. (now Shire Biochem, Inc.); PCT/US02/01531 (WO 02/057425;filed Jan. 18, 2002); PCT/US02/03086 (WO 02/057287; filed Jan. 18,2002); U.S. Pat. Nos. 7,202,224; 7,125,855; 7,105,499; and 6,777,395 byMerck & Co., Inc.; PCT/EP01/09633 (WO 02/18404; published Aug. 21,2001); US 2006/0040890; 2005/0038240; 2004/0121980; 6,846,810;6,784,166; and 6,660,721 by Roche; PCT Publication Nos. WO 01/79246(filed Apr. 13, 2001), WO 02/32920 (filed Oct. 18, 2001) and WO02/48165; US 2005/0009737; US 2005/0009737; 7,094,770; and 6,927,291 byPharmasset, Ltd.

Further compounds that can be used as second agents to treat hepatitis Cvirus are disclosed in PCT Publication No. WO 99/43691 to EmoryUniversity, entitled “2′-Fluoronucleosides”. The use of certain2′-fluoronucleosides to treat HCV is disclosed.

Other compounds that can be used as second agents include1-amino-alkylcyclohexanes (U.S. Pat. No. 6,034,134 to Gold et al.),alkyl lipids (U.S. Pat. No. 5,922,757 to Chojkier et al.), vitamin E andother antioxidants (U.S. Pat. No. 5,922,757 to Chojkier et al.),squalene, amantadine, bile acids (U.S. Pat. No. 5,846,964 to Ozeki etal.), N-(phosphonoacetyl)-L-aspartic acid, (U.S. Pat. No. 5,830,905 toDiana et al.), benzenedicarboxamides (U.S. Pat. No. 5,633,388 to Dianaet al.), polyadenylic acid derivatives (U.S. Pat. No. 5,496,546 to Wanget al.), 2′,3′-dideoxyinosine (U.S. Pat. No. 5,026,687 to Yarchoan etal.), benzimidazoles (U.S. Pat. No. 5,891,874 to Colacino et al.), plantextracts (U.S. Pat. No. 5,837,257 to Tsai et al., U.S. Pat. No.5,725,859 to Omer et al., and U.S. Pat. No. 6,056,961), and piperidines(U.S. Pat. No. 5,830,905 to Diana et al.).

In certain embodiments, a compound of Formula 2001, I-XXIX, 1-23byii and1001-1002, or a composition comprising a compound of Formula 2001,I-XXIX, 1-23byii and 1001-1002, is administered in combination oralternation with a second anti-viral agent, wherein the secondanti-viral agent is an interferon, a nucleotide analogue, a polymeraseinhibitor, an NS3 protease inhibitor, an NS5A inhibitor, an entryinhibitor, a non-nucleoside polymerase inhibitor, a cyclosporine immuneinhibitor, an NS4A antagonist, an NS4B-RNA binding inhibitor, a lockednucleic acid mRNA inhibitor, a cyclophilin inhibitor, or combinationthereof.

Exemplary Second Therapeutic Agents for Treatment of HCV

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus interferon, such as Intron A® (interferon alfa-2b) and; Roferon A®(Recombinant interferon alfa-2a), Infergen® (consensus interferon;interferon alfacon-1), PEG-Intron® (pegylated interferon alfa-2b), andPegasys® (pegylated interferon alfa-2a). In certain embodiments, one ormore compounds provided herein can be administered in combination oralternation with ribavirin and in combination or alternation with ananti-hepatitis C virus interferon. In certain embodiments, one or morecompounds provided herein can be administered in combination oralternation with ribavirin, in combination or alternation with ananti-hepatitis C virus interferon, and in combination or alternationwith an anti-hepatitis C virus protease inhibitor. In certainembodiments, one or more compounds provided herein can be administeredin combination or alternation with ribavirin. In certain embodiments,one or more compounds provided herein can be administered in combinationor alternation with an anti-hepatitis C virus interferon and withoutribavirin. In certain embodiments, one or more compounds provided hereincan be administered in combination or alternation with an anti-hepatitisC virus interferon, in combination or alternation with an anti-hepatitisC virus protease inhibitor, and without ribavirin.

In certain embodiments, the anti-hepatitis C virus interferon isinfergen, IL-29 (PEGInterferon lambda), R7025 (Maxy-alpha), Belerofon,Oral Interferon alpha, BLX-883 (Locteron), omega interferon, multiferon,medusa interferon, Albuferon or REBIF®.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus polymerase inhibitor, such as ribavirin, viramidine, HCV POL, NM283 (valopicitabine), MK-0608, 7-Fluoro-MK-0608, PSI-6130, R1626,PSI-6206, PSI-938, R1479, HCV-796, VX-950 (Telaprevir, Vertex), GS 9190NN (Gilead), GS 9256 (Gilead), PSI-7792 (BMS), BI 207127 (BI), R7128(Roche), PSI-7977 (sofosbuvir, Sovaldi; Pharmasset), PSI-938(Pharmasset), VX-222 (Vertex), ALS-2200 (Vertex), ALS-2158 (Vertex),MK-0608 (Merck), TMC649128 (Medivir), PF-868554 (Pfizer), PF-4878691(Pfizer), ANA598 (Roche), VCH-759 (Vertex), IDX184 (Idenix), IDX375(Idenix), A-837093 (Abbott), GS 9190 (Gilead), GSK625433(GlaxoSmithKline), ABT-072 (Abbott), ABT-333 (Abbott), INX-189(Inhibitex), or EDP-239 (Enanta).

In certain embodiments, the one or more compounds provided herein can beadministered in combination with ribavarin and an anti-hepatitis C virusinterferon, such as Intron A® (interferon alfa-2b) and Pegasys®(Peginterferon alfa-2a); Roferon A® (Recombinant interferon alfa-2a),Infergen® (consensus interferon; interferon alfacon-1), PEG-Intron®(pegylated interferon alfa-2b), Zalbin (albinterferon alfa-2b), omegainterferon, pegylated interferon lambda, and Pegasys® (pegylatedinterferon alfa-2a).

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus protease inhibitor such as ITMN-191, SCH 503034 (boceprevir),VX950 (telaprevir), VX985, VX500, VX813, PHX1766, BMS-650032, GS 9256,BI 201335, IDX320, R7227, MK-7009 (vaniprevir), simeprevir (TMC435),BMS-791325, ACH-1625, ACH-2684, ABT-450, or AVL-181.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an HCV NS5A inhibitor,such as BMS-790052 (daclatasvir, Bristol-Myers Squibb), PPI-461(Presidio Pharmaceuticals), PPI-1301 (Presidio Pharmaceuticals), IDX-719(samatasvir, Idenix Pharmaceuticals), AZD7295 (Arrow Therapeutics,AstraZeneca), EDP-239 (Enanta), ACH-2928 (Achillion), ACH-3102(Achillion), ABT-267 (Abbott), or GS-5885 (Gilead).

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus vaccine, such as TG4040, PeviPRO™, CGI-5005, HCV/MF59, GV1001,IC41, GNI-103, GenPhar HCV vaccine, C-Vaxin, CSL123, Hepavaxx C,ChronVac-C® or INNO0101 (E1).

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus monoclonal antibody, such as MBL-HCV1, AB68 or XTL-6865 (formerlyHepX-C); or an anti-hepatitis C virus polyclonal antibody, such ascicavir.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an anti-hepatitis Cvirus immunomodulator, such as Zadaxin® (thymalfasin), SCV-07, NOV-205,or Oglufanide.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with cyclophilin inhibitor,such as Enanta cyclophilin binder, SCY-635, or Debio-025.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with Nexavar, doxorubicin,PI-88, amantadine, JBK-122, VGX-410C, MX-3253 (Ceglosivir), Suvus(BIVN-401 or virostat), PF-03491390 (formerly IDN-6556), G126270,UT-231B, DEBIO-025, EMZ702, ACH-0137171, MitoQ, ANA975, AVI-4065,Bavituxinab (Tarvacin), Alinia (nitrazoxanide) or PYN17.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with simeprevir, sofosbuvir,samatasvir, telaprevir, boceprevir, interferon alfacon-1, interferonalfa-2b, pegylated interferon alpha 2a, pegylated interferon alpha 2b,ribavirin, or combinations thereof.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with a protease inhibitor. Incertain embodiments, one or more compounds provided herein can beadministered in combination or alternation with telaprevir. In certainembodiments, one or more compounds provided herein can be administeredin combination or alternation with boceprevir.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with a protease inhibitor andin combination or alternation with ribavirin. In certain embodiments,one or more compounds provided herein can be administered in combinationor alternation with telaprevir and in combination or alternation withribavirin. In certain embodiments, one or more compounds provided hereincan be administered in combination or alternation with boceprevir and incombination or alternation with ribavirin.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with a protease inhibitor andnot in combination or alternation with ribavirin. In certainembodiments, one or more compounds provided herein can be administeredin combination or alternation with telaprevir and not in combination oralternation with ribavirin. In certain embodiments, one or morecompounds provided herein can be administered in combination oralternation with boceprevir and not in combination or alternation withribavirin.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an interferon. Incertain embodiments, one or more compounds provided herein can beadministered in combination or alternation with interferon alfacon-1. Incertain embodiments, one or more compounds provided herein can beadministered in combination or alternation with interferon alfa-2b. Incertain embodiments, one or more compounds provided herein can beadministered in combination or alternation with pegylated interferonalpha 2a. In certain embodiments, one or more compounds provided hereincan be administered in combination or alternation with pegylatedinterferon alpha 2b.

In certain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an interferon and incombination or alternation with ribavirin. In certain embodiments, oneor more compounds provided herein can be administered in combination oralternation with interferon alfacon-1 and in combination or alternationwith ribavirin. In certain embodiments, one or more compounds providedherein can be administered in combination or alternation with interferonalfa-2b and in combination or alternation with ribavirin. In certainembodiments, one or more compounds provided herein can be administeredin combination or alternation with pegylated interferon alpha 2a and incombination or alternation with ribavirin. In certain embodiments, oneor more compounds provided herein can be administered in combination oralternation with pegylated interferon alpha 2b and in combination oralternation with ribavirin.

In certain embodiments, one or more compounds can be administered incombination or alternation with one or more of the second agentsprovided herein and not in combination or alternation with ribavirin. Incertain embodiments, one or more compounds provided herein can beadministered in combination or alternation with an interferon and not incombination or alternation with ribavirin. In certain embodiments, oneor more compounds provided herein can be administered in combination oralternation with interferon alfacon-1 and not in combination oralternation with ribavirin. In certain embodiments, one or morecompounds provided herein can be administered in combination oralternation with interferon alfa-2b and not in combination oralternation with ribavirin. In certain embodiments, one or morecompounds provided herein can be administered in combination oralternation with pegylated interferon alpha 2a and not in combination oralternation with ribavirin. In certain embodiments, one or morecompounds provided herein can be administered in combination oralternation with pegylated interferon alpha 2b and not in combination oralternation with ribavirin.

EXAMPLES

As used herein, the symbols and conventions used in these processes,schemes and examples, regardless of whether a particular abbreviation isspecifically defined, are consistent with those used in the contemporaryscientific literature, for example, the Journal of the American ChemicalSociety or the Journal of Biological Chemistry. Specifically, butwithout limitation, the following abbreviations may be used in theexamples and throughout the specification: g (grams); mg (milligrams);mL (milliliters); μL (microliters); mM (millimolar); μM (micromolar); Hz(Hertz); MHz (megahertz); mmol (millimoles); hr or hrs (hours); min(minutes); MS (mass spectrometry); ESI (electrospray ionization); TLC(thin layer chromatography); HPLC (high pressure liquid chromatography);THF (tetrahydrofuran); CDCl₃ (deuterated chloroform); AcOH (aceticacid); DCM (dichloromethane); DMSO (dimethylsulfoxide); DMSO-d₆(deuterated dimethylsulfoxide); EtOAc (ethyl acetate); MeOH (methanol);and BOC (t-butyloxycarbonyl).

For all of the following examples, standard work-up and purificationmethods known to those skilled in the art can be utilized. Unlessotherwise indicated, all temperatures are expressed in ° C. (degreesCentigrade). All reactions are conducted at room temperature unlessotherwise noted. Synthetic methodologies illustrated herein are intendedto exemplify the applicable chemistry through the use of specificexamples and are not indicative of the scope of the disclosure.

Example 1 Preparation of Thiophosphate Nucleosides Compounds 8b, 9b and10b

General Method A.

The following procedure was used to obtain compounds 8b and 9b.

To a solution of 2′-chloro-2′-C-methyl-uridine (3.61 mmol) in triethylphosphate (5 mL) at 0° C. under nitrogen was added POCl₃ (5.42 mmol).The reaction mixture was allowed to warm to room temperature and stirredat this temperature during 12 hours. The reaction mixture was cooleddown again to 0° C. and L-Alanine isopropyl ester hydrochloride (3.98mmol) was added followed by addition of triethylamine (18.07 mmol). Thereaction was stirred at room temperature during 20 minutes.

The reaction mixture was cooled down to 0° C. and appropriate thiol(14.46 mmol) in CH₃CN (4 mL) was added followed by addition of DBU(14.46 mmol). The reaction mixture was stirred at room temperatureduring 1 hour. The mixture was diluted with ethyl acetate and washedwith brine. The organic layer was dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The crude was purified bychromatography on a Biotage column (eluent: CH₂Cl₂ to CH₂Cl₂/CH₃OH 8/2)and by preparative HPLC to give the pure diastereoisomers.

Compound 8b

Compound 8b Dia 1

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.25 (d, J=6.07 Hz, 3H),1.27 (d, J=6.07 Hz, 3H), 1.41 (d, J=7.31 Hz, 3H), 1.54 (s, 3H),1.62-1.78 (m, 6H), 2.11-2.18 (m, 2H), 3.48-3.56 (m, 1H), 3.87-3.95 (m,1H), 3.99-4.02 (m, 1H), 4.18-4.22 (m, 1H), 4.36 (ddd, J=11.97 Hz, J=5.75Hz, J=2.64 Hz 1H), 4.54 (ddd, J=12.08 Hz, J=5.83 Hz, J=2.12 Hz, 1H),5.01 (heptuplet, J=6.25 Hz, 1H), 5.80 (d, J=8.16 Hz, 1H), 6.40 (s, 1H),7.85 (d, J=8.16 Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 33.78 (s,1P); MS (ESI) m/z=554.2 (MH⁺).

Compound 8b Dia 2

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.26 (d, J=6.21 Hz, 3H),1.27 (d, J=6.21 Hz, 3H), 1.42 (d, J=7.14 Hz, 3H), 1.54 (s, 3H),1.58-1.78 (m, 6H), 2.07-2.18 (m, 2H), 3.47-3.54 (m, 1H), 3.92-4.00 (m,1H), 4.03-4.06 (m, 1H), 4.17-4.21 (m, 1H), 4.32 (ddd, J=11.98 Hz, J=5.75Hz, J=2.68 Hz 1H), 4.50 (ddd, J=11.98 Hz, J=6.23 Hz, J=1.97 Hz, 1H),5.02 (heptuplet, J=6.21 Hz, 1H), 5.82 (d, J=8.16 Hz, 1H), 6.40 (s, 1H),7.82 (d, J=8.16 Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 34.78 (s,1P); MS (ESI) m/z=554.2 (MH⁺).

Compound 9b

Diastereoisomer 1

white solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.21 (d, J=7.20 Hz, 3H),1.24 (d, J=6.22 Hz, 3H), 1.26 (d, J=6.22 Hz, 3H), 1.52 (s, 3H),3.80-3.88 (m, 1H), 3.92-3.94 (m, 1H), 4.18-4.22 (m, 1H), 4.41-4.46 (m,1H), 4.60-4.65 (m, 1H), 4.99 (heptuplet, J=6.20 Hz, 1H), 5.79 (d, J=8.16Hz, 1H), 6.40 (s, 1H), 7.37-7.44 (m, 3H), 7.62-7.65 (m, 2H), 7.77 (d,J=8.16 Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 30.45 (s, 1P); MS(ESI) m/z=562 (MH⁺).

Diastereoisomer 2

white solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.25 (d, J=6.20 Hz, 3H),1.28 (d, J=6.20 Hz, 3H), 1.36 (d, J=7.07 Hz, 3H), 1.52 (s, 3H),3.91-4.02 (m, 2H), 4.16-4.20 (m, 1H), 4.36-4.41 (m, 1H), 4.56-4.61 (m,1H), 4.99 (heptuplet, J=6.23 Hz, 1H), 5.64 (d, J=8.13 Hz, 1H), 6.39 (s,1H), 7.32-7.40 (m, 3H), 7.58-7.62 (m, 3H); ³¹P NMR (MeOD, 161.98 MHz) δ(ppm) 31 (s, 1P); MS (ESI) m/z=562 (MH⁺).

Compound 10b

To a solution of 2′-C-methyl-6-ethoxy-guanosine (3.07 mmol) in triethylphosphate (3 mL) at 0° C. under nitrogen was added1-dichlorophosphorylsulfanylethane (Pestic. Sci. (1984), 15, 553-556)(6.15 mmol). The reaction mixture was stirred at room temperature. To asolution of L-Alanine ethyl ester hydrochloride (4.61 mmol) in CH₃CN (5mL) was added dropwise triethylamine (15.37 mmol) and the reactionmixture was stirred at 0° C. during 30 minutes before addition dropwiseto the first reaction mixture (pH must be neutral). After a night atroom temperature, the reaction mixture was quenched on phosphate buffersolution and extracted with ethyl acetate. The organic layer was driedover Na₂SO₄, filtered, and concentrated under reduced pressure. Thecrude was purified by chromatography on a silica gel (eluent:CH₂Cl₂/CH₃OH 0 to 10%) and by preparative HPLC to give a mixture ofdiastereoisomers.

¹H NMR (DMSO-d₆, 400 MHz) δ (ppm) 0.796 (s, 1.5H), 0.803 (s, 1.5H),1.14-1.22 (m, 6H), 1.28 (d, J=7.09 Hz, 3H), 1.35 (t, J=7.09 Hz, 3H),2.65-2.77 (m, 2H), 3.72-3.88 (m, 1H), 4.00-4.10 (m, 4H), 4.17-4.36 (m,2H), 4.44 (q, J=7.07 Hz, 2H), 5.21 (d, J=3.18 Hz, 1H), 5.40 (dd, J=6.13Hz and 15.46 Hz, 1H), 5.837 (s, 0.5H), 5.841 (s, 0.5H), 6.11-6.18 (m,1H), 6.46 (s, 2H), 7.91 (s, 0.5H), 7.92 (s, 0.5H); ³¹P NMR (MeOD, 161.98MHz) δ (ppm) 31.32 (s, 0.5P), 31.78 (s, 0.5P); MS (ESI) m/z=549.2 (MH⁺).

Compound 11b was synthesized from2′-fluoro-2′-C-methyl-6-ethoxy-guanosine according to the proceduredescribed for compound 10b in Scheme 2. The mixture of diastereoisomerswas separated by MS-preparative HPLC.

Compound 11b Dia 1

White solid; ¹H NMR (DMSO-d₆, 400 MHz) δ (ppm) 1.07 (d, J=22.13 Hz, 3H),1.15-1.20 (m, 6H), 1.27 (d, J=7.01 Hz, 3H), 1.34 (t, J=7.01 Hz, 3H),2.67-2.74 (m, 2H), 3.74-3.83 (m, 1H), 4.05-4.11 (m, 3H), 4.24-4.39 (m,3H), 4.44 (q, J=7.01 Hz, 2H), 5.85 (brs, 1H), 6.09 (d, J=19.04 Hz, 1H),6.19 (dd, J=13.03 Hz, 10.02 Hz, 1H), 6.57 (brs, 2H), 7.94 (s, 1H); ³¹PNMR (DMSO-d₆, 161.98 MHz) δ (ppm) 31.44 (s, 1P); MS (ESI) m/z=551.3(MH⁺).

Compound 11b Dia 2

White solid; ¹H NMR (DMSO-d₆, 400 MHz) δ (ppm) 1.07 (d, J=22.73 Hz, 3H),1.14-1.21 (m, 6H), 1.27 (d, J=7.05 Hz, 3H), 1.35 (t, J=7.05 Hz, 3H),2.69-2.77 (m, 2H), 3.78-3.85 (m, 1H), 4.03-4.08 (m, 3H), 4.19-4.38 (m,3H), 4.44 (q, J=6.84 Hz, 2H), 5.82 (brs, 1H), 6.08 (d, J=19.22 Hz, 1H),6.14-6.20 (m, 1H), 6.57 (brs, 2H), 7.95 (s, 1H); ³¹P NMR (DMSO-d₆,161.98 MHz) δ (ppm) 31.94 (s, 1P); MS (ESI) m/z=551.3 (MH⁺).

Compound 12b was synthesized from 2′-C-methyl-uridine and N-propanethiolaccording to procedure described for compound 8b in Scheme 1 withtriethylamine instead of DBU. The mixture of diastereoisomers wasseparated by MS-preparative HPLC.

Compound 12b Dia 1

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 0.91 (t, J=7.26 Hz, 3H),1.07 (s, 3H), 1.16 (t, J=5.76 Hz, 6H), 1.26-1.31 (m, 3H), 1.58-1.67 (m,2H), 2.69-2.77 (m, 2H), 3.71 (d, J=9.25 Hz, 1H), 3.75-3.83 (m, 1H),3.98-4.02 (m, 1H), 4.23 (ddd, J=11.83 Hz, J=5.92 Hz, J=2.90 Hz, 1H),4.40 (ddd, J=11.71 Hz, J=5.86 Hz, J=2.05 Hz, 1H), 4.91 (heptuplet,J=6.31 Hz, 1H), 5.68 (d, J=8.11 Hz, 1H), 5.88 (s, 1H), 7.67 (d, J=8.11Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 34.88 (s, 1P); MS (ESI)m/z=510.2 (MH⁺).

Compound 12b Dia 2

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 0.88 (t, J=7.36 Hz, 3H),1.07 (s, 3H), 1.16 (d, J=6.20 Hz, 3H), 1.17 (d, J=6.20 Hz, 3H), 1.31 (d,J=7.11 Hz, 3H), 1.55-1.63 (m, 2H), 2.68-2.75 (m, 2H), 3.73 (d, J=9.30Hz, 1H), 3.80-3.88 (m, 1H), 3.97-4.01 (m, 1H), 4.20 (ddd, J=11.81 Hz,J=5.91 Hz, J=3.21 Hz, 1H), 4.36 (ddd, J=11.81 Hz, J=6.71 Hz, J=2.05 Hz,1H), 4.92 (heptuplet, J=6.27 Hz, 1H), 5.70 (d, J=8.09 Hz, 1H), 5.88 (s,1H), 7.65 (d, J=8.09 Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 35.64(s, 1P); MS (ESI) m/z=510.2 (MH⁺).

Compound 13b was synthesized from 2′-C-methyl-guanosine according toprocedure described for compound 10b in Scheme 2.

Compound 13b

White solid; ¹H NMR (DMSO-d₆, 400 MHz) δ (ppm) 0.82 (s, 3H), 1.14-1.22(m, 6H), 1.28 (d, J=7.06 Hz, 3H), 2.68-2.77 (m, 2H), 3.77-3.88 (m, 1H),3.95-4.11 (m, 4H), 4.15-4.32 (m, 2H), 5.17 (s, 0.85H), 5.18 (s, 0.15H),5.40 (d, J=5.86 Hz, 0.85H), 5.43 (d, J=6.55 Hz, 0.15H), 5.74 (s, 1H),6.16 (dd, J=13.10 Hz, 10.34 Hz, 1H), 6.52 (brs, 2H), 7.745 (s, 015H),7.755 (s, 085H); ³¹P NMR (DMSO-d₆, 161.98 MHz) δ (ppm) 31.81 (s, 0.85),31.33 (s, 0.15P); MS (ESI) m/z=521.2 (MH⁺).

Compound 14b was synthesized from 2′-chloro-2′-C-methyl-uridine andN-propanethiol according to procedure described for compound 8b inScheme 1 with triethylamine instead of DBU. The mixture ofdiastereoisomers was separated by MS-preparative HPLC.

Compound 14b Dia 1

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 0.91 (t, J=7.28 Hz, 3H),11.15-1.18 (m, 6H), 1.31 (d, J=7.33 Hz, 3H), 1.44 (s, 3H), 2.71-2.78 (m,2H), 3.78-3.82 (m, 1H), 3.89-3.91 (m, 1H), 4.07-4.16 (m, 4H), 4.23-4.28(m, 1H), 4.41-4.48 (m, 2H), 4.91 (heptuplet, J=6.23 Hz, 1H), 5.71 (d,J=8.19 Hz, 1H), 6.30 (brs, 1H), 7.73 (d, J=8.19 Hz, 1H); ³¹P NMR (MeOD,161.98 MHz) δ (ppm) 34.89 (s, 1P); MS (ESI) m/z=528.2 (MH⁺).

Compound 14b Dia 2

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 0.89 (t, J=7.30 Hz, 3H),1.16 (d, J=6.20 Hz, 3H), 1.17 (d, J=6.28 Hz, 3H), 1.32 (d, J=7.11 Hz,3H), 1.44 (s, 3H), 2.69-2.77 (m, 2H), 3.83-3.95 (m, 2H), 4.07-4.13 (m,2H), 4.20-4.25 (m, 1H), 4.37-4.50 (m, 2H), 4.92 (heptuplet, J=6.23 Hz,1H), 5.72 (d, J=8.11 Hz, 1H), 6.31 (brs, 1H), 7.71 (d, J=8.11 Hz, 1H);³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 35.75 (s, 1P); MS (ESI) m/z=528.2(MH⁺).

Compound 17b was synthesized from 2′-chloro-2′-C-methyl-uridine andmethyl thioglycolate according to procedure described for compound 8b inScheme 1.

Compound 17b

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.25-1.30 (m, 6H), 1.42 (t,J=7.45 Hz, 3H), 1.56 (s, 3H), 3.72-3.74 (m, 3H), 3.88-4.10 (m, 2H),4.18-4.22 (m, 1H), 4.33-4.43 (m, 1H), 4.54-4.60 (m, 1H), 5.02(heptuplet, J=6.20 Hz, 1H), 5.83 (d, J=8.15 Hz, 0.55H), 5.84 (d, J=8.15Hz, 0.45H), 6.43 (brs, 1H), 7.79 (d, J=8.15 Hz, 0.45H), 7.84 (d, J=8.15Hz, 0.55H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 33.44 (s, 0.45P), 32.57(s, 0.55P); MS (ESI) m/z=558.2 (MH⁺).

Compound 1b Two Separated Diastereomers at Phosphorus

The chiral reagent A-2 was prepared by the following scheme; 97% ee wasobserved.

A-2, (S)-1-(cyclopentyloxy)-3-(2-mercaptophenyl)-1-oxopropan-2-aminiumchloride: LCMS (ES⁺, m/e=266.2, M⁺+1), ¹H NMR (300 MHz, D₂O) δ 7.33 (m.1H), 7.13 (m, 3H), 5.08 (m, 1H), 4.32 (t, 1H), 3.17 (m, 2H), 1.42-1.71(m, 8H).

The nucleoside (A-N1, 0.44 g, 1.42 mmol) was stirred intriethylphosphate (PO(OEt)₃, 20 mL) and was cooled to −20° C.Phosphorous oxychloride (POCl₃, 0.28 g, 1.85 mmol, 1.3 eq) was addeddrop-wise and the mixture was stirred at −20° C. for 30 min. The mixturewas allowed to warm to 0° C. and was stirred at this temperature for 4hours. This mixture was then cooled to −50° C. and the aminehydrochloride (A-2, 0.43 g, 1.42 mmol, 1.0 eq) was added followed bytriethylamine (TEA, 0.59 mL, 4.6 mmol, 2.3 eq). After stirring at −50°C. for 30 minutes, an additional portion of TEA (1.11 mL, 6.13 mmol, 4.3eq) was added and the mixture was allowed to warm to room temperatureand was stirred overnight. The crude product was precipitated by pouringthe reaction mixture into petroleum ether (300 mL) and was isolated byvacuum filtration. This material (1.58 g) was purified by preparativeHPLC affording compound 1b, diastereomer 1 (33 mg, 37%) and compound 1b,diastereomer 2 (20 mg, 23%).

Diastereomer 1

HPLC (Method 1), RT 13.55 min, 98.4 A %; LCMS, (ES⁺, m/e=621.1, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.92 (s, 1H), 7.47 (d, 1H), 7.29 (m, 1H), 7.18(m, 2H), 5.94 (s, 1H), 5.14 (m, 1H), 4.53-4.83 (m, 3H), 4.33 (m, 1H),4.20 (m, 2H), 4.06 (s, 3H), 3.68 (m, 1H), 3.40 (m, 1H), 1.64-1.89 (m,8H), 0.96 (s, 3H); ³¹P NMR (121 MHz, CD₃OD) δ 28.361.

Diastereomer 2

HPLC (Method 1), RT 17.45 min, 98.9 A %; LCMS, (ES⁺, m/e=621.1, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.85 (s, 1H), 7.54 (d, 1H), 7.20 (m, 3H), 5.93(s, 1H), 5.11 (m, 1H), 4.62 (m, 1H), 4.40 (m, 1H), 4.18 (m, 3H), 4.05(s, 3H), 3.57 (m, 1H), 3.37 (m, 1H), 1.64-1.89 (m, 8H), 0.91 (s, 3H);³¹P NMR (121 MHz, CD₃OD) δ 27.984.

HPLC Method 1:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 45%-45%-97% MeCN(0-30-35 min), 250 nm.

Compound 2by Mixture of Diastereomers at Phosphorus

B-3 was prepared according to the following scheme:

L-Cystine (10.0 g, 41.6 mmol) was dissolved in a solution of sodiumcarbonate (9.266 g, 87.3 mmol, 2.1 eq) in water (400 mL). The mixturewas stirred vigorously and a solution of methyl chloroformate (8.22 g,87.0 mmol, 2.1 eq) in THF (160 mL) was added drop-wise at roomtemperature. The mixture was stirred for 16 hours and aqueous HCl wasadded to adjust the pH to between 1 and 2. This mixture was thenextracted with DCM (3×200 mL). The combined organic extracts were driedover sodium sulfate and were concentrated under reduced pressureproviding 9.5 g (65%) of BI-1 as a colorless syrup.

BI-1 (14.8 g, 41.5 mmol) was added to DCM (350 mL).Dimethylaminopyridine (DMAP, 8.1 g, 66.4 mmol, and 1.6 eq) and isopropylalcohol (14.4 g, 240 mmol, 6 eq) were added and the mixture was stirredand cooled to 0° C. A solution of dicyclohexylcarbodiimide (DCC, 18.8 g,91.3 mmol, 2.2 eq) dissolved in DCM (150 mL) was added drop-wise. Uponcompletion of the addition, the ice-bath was removed and the mixturewarmed to room temperature and stirred overnight. The resultingsuspension was filtered and the filtrate was concentrated under reducedpressure. The residue was purified by flash column-chromatography(silica gel, 25% EtOAc/Petroleum Ether) affording 11.5 g (62%) of BI-2as a white-colored solid.

BI-2 (5.0 g, 11.3 mmol, 1.0 eq) was dissolved in methanol (8 mL). Asolution of acetic acid (0.2 mL) and sodium acetate (300 mg) in water (4mL) were added followed by triphenylphosphine (Ph₃P, 5.0 g, 19.1 mmol,1.7 eq). The resulting suspension was stirred at 80° C. overnight. Themixture was cooled to room temperature and was extracted with DCM (3×15mL). The combined organic phases were dried over sodium sulfate and wereconcentrated under reduced pressure. Purification of the crude productby flash column-chromatography (silica gel) afforded 4.5 g (90%) of B-3as a colorless oil.

B-2 was prepared as described below.

L-Alanine (10 g, 112.4 mmol, 1 eq) was dissolved in isopropanol (150mL). Thionyl chloride (25 mL, 337 mmol, 3 eq) was added and the mixturewas heated to reflux (100° C.) until the mixture became clear. Themixture was cooled and was concentrated under reduced pressure. Theresulting material was crystallized to provide B-2 as a white-coloredsolid.

The nucleoside (B-N1, 933 mg, 3 mmol, 1 eq) was dissolved intriethylphosphate (10 mL). The mixture was cooled to 0° C. (ice-bath)under a nitrogen atmosphere and phosphorous oxychloride (504 mg, 3.3mmol, 1.1 eq) was added drop-wise. The mixture was stirred at 0° C. for1 hour. A solution of B-2 (501 mg, 3 mmol, 1.0 eq) and triethylamine(637 mg, 6.3 mmol, 2.1 eq) in triethylphosphate (2 mL) was prepared andwas added drop-wise to the solution of B-N1. The mixture was allowed tostir at 0° C. for 30 minutes after which time a solution of B-3 (732 mg,3.3 mmol, 1.1 eq) and triethylamine (1.183 g, 11.7 mmol, 3.9 eq) in DCM(5 mL) was added. The ice-bath was removed and the mixture was allowedto warm to room temperature and stirred overnight. The mixture waspoured into 200 mL of petroleum ether and the resulting precipitate wascollected by vacuum filtration. The crude product was purified by flashcolumn-chromatography (4% MeOH/DCM) affording 500 mg of semi-pure 2by.This material was purified further by preparative HPLC to yield a 400 mgsample of the pure mixture of isomers 2by.

2by

HPLC (Method 2), RT 4.794 min, 99.4 A %; LCMS (ES⁺, m/e=708, M⁺+1); ¹HNMR (300 MHz, CD₃OD) δ 7.96, 7.96 (2×s, 1H), 5.98, 5.97 (2×s, 1H), 4.90(m, 2H), 4.48 (m, 2H), 4.22 (m, 3H), 4.05 (s, 3H), 3.89 (1H, m), 3.65(s, 3H), 3.35 (s, 2H), 3.31 (m, 1H), 1.33 (m, 3H), 1.20 (m, 12H), 0.976,0.965 (2×s, 3H); ³¹P NMR (121 MHz, CD₃OD) δ 35.67, 35.50. HPLC Method 2:C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 25%-97% MeCN (0-15min), 250 nm.

Compound 23b Two Diastereomers Isolated From a Four Isomer Mixture

B-4 and B-2 were prepared as follows.

BI-3 (15 g, 124 mmol, 1 eq) and potassium cyanate (11.05 g, 136 mmol,1.2 eq) were dissolved in water (75 mL). The mixture was heated toreflux and aqueous HCl (10%, 50 mL) was added. After refluxing for 30minutes, the mixture was cooled to room temperature. DCM was added. Thematerial which precipitated was isolated by filtration. This materialwas the desired product and was of sufficient purity to use in thecoupling step.

L Alanine (10 g, 112.4 mmol, 1 eq) was dissolved in isopropanol (150mL). Thionyl chloride (25 mL, 337 mmol, 3 eq) was added and the mixturewas heated to reflux (100° C.) until the mixture became clear. Themixture was cooled and was concentrated under reduced pressure. Theresulting material was crystallized from ether and petroleum ether toprovide B-2 a white-colored solid.

The nucleoside (B-N1, 1.5 g, 4.82 mmol) was stirred in triethylphosphate(PO(OEt)₃, 30 mL) and was cooled to −20° C. Phosphorous oxychloride(POCl₃, 0.96 g, 6.26 mmol, 1.3 eq) was added drop-wise and the mixturewas stirred at −20° C. for 30 min. The mixture was allowed to warm to 0°C. and was stirred at this temperature for 4 hours. This mixture wascooled to −50° C. (dry ice/EtOH) B-2 (0.724 g, 4.34 mmol, 0.9 eq) wasadded followed by triethylamine (TEA, 1.54 mL, 11.08 mmol, 2.3 eq).After stirring at −50° C. for 30 minutes, B-4 (2.11 g, 14.46 mmol, 3 eq)was added to the mixture followed by triethylamine (TEA, 2.88 mL, 20.72mmol, 4.3 eq). The mixture was allowed to warm to room temperature andwas stirred overnight. Petroleum ether was added to the mixture and theprecipitated material was isolated by vacuum filtration. This materialwas purified by column chromatography (silica gel, 200-300 mesh) usingMeOH/DCM (0→7%) providing the crude product (570 mg). The crude productpurified by preparative column chromatography affording compound 23b,diastereomer 1 (10 mg, 0.33%) and compound 23b, diastereomer 2 (10 mg,0.33%).

Compound 23b, Diastereomer 1

HPLC (Method 3), RT 15.23 min, 96.9 A %; LCMS (ES⁺, m/e=633.1, M⁺+1); ¹HNMR (300 MHz, CD₃OD) δ 7.95 (s. 1H), 5.96 (s, 1H), 4.97 (m, 1H), 4.48(m, 3H), 4.35 (m, 1H), 4.18 (m, 1H), 4.05 (s, 3H), 4.02 (m, 3H), 3.80(m, 1H), 3.60 (m, 1H), 1.34 (m, 3H), 1.22 (m, 6H), 0.98 (s, 3H); ³¹P NMR(121 MHz, CD₃OD) δ 7.40.

Compound 23b, Diastereomer 2

HPLC (Method 3), RT 16.58 min, 98.60 A %; LCMS (ES⁺, m/e=633.1, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.97 (s, 1H), 5.98 (s, 1H), 4.97 (m, 1H), 4.47(m, 3H), 4.35 (m, 1H), 4.18 (m, 1H), 4.06 (s, 3H), 4.02 (s, 3H), 3.80(m, 1H), 3.60 (m, 1H), 1.37 (m, 3H), 1.27 (m, 6H), 0.98 (s, 3H); ³¹P NMR(121 MHz, CD₃OD) δ 7.14.

HPLC Method 3:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 15%-15%-50%-97% MeCN(0-20-25-30 min), 282 nm.

Compound 3b Two Diastereomers Isolated From a Four Isomer Mixture

The nucleoside (B-N2, 1.5 g, 4.82 mmol) was stirred in triethylphosphate(PO(OEt)₃, 30 mL) and was cooled to −20° C. Phosphorous oxychloride(POCl₃, 0.96 g, 6.26 mmol, 1.3 eq) was added drop-wise and the mixturewas stirred at −20° C. for 30 min. The mixture was allowed to warm to 0°C. and was stirred at this temperature for 4 hours. This mixture wascooled to −50° C. (dry ice/EtOH) B-2 (0.724 g, 4.34 mmol, 0.9 eq) wasadded followed by triethylamine (TEA, 1.54 mL, 11.08 mmol, 2.3 eq).After stirring at −50° C. for 30 minutes, B-4 (2.11 g, 14.46 mmol, 3 eq)was added and the mixture followed by triethylamine (TEA, 2.88 mL, 20.72mmol, 4.3 eq). The mixture was allowed to warm to room temperature andwas stirred overnight. Petroleum ether was added to the mixture and theprecipitated material was isolated by vacuum filtration. This materialwas purified by column chromatography (silica gel, 200-300 mesh) usingMeOH/DCM (0→7%) providing the crude product (570 mg). The crude productwas purified by preparative column chromatography affording compound 3b,diastereomer 1 (10 mg, 0.33%) and compound 3b, diastereomer 2 (10 mg,0.33%).

Compound 3b, Diastereomer 1

HPLC (Method 4), RT 27.68 min, 97.5 A %; LCMS (ES⁺, m/e=649.2, M⁺+1); ¹HNMR (300 MHz, CD₃OD) δ 7.94 (s, 1H), 6.16 (d, 1H), 4.96 (m, 1H), 4.47(m, 6H), 4.20 (m, 1H), 4.00 (m, 1H), 3.78 (m, 1H), 3.56 (m, 1H), 3.32(d, 3H), 1.43 (t, 3H), 1.35 (m, 3H), 1.20 (m, 6H); ³¹P NMR (121 MHz,CD₃OD) δ 7.39.

Compound 3b, Diastereomer 2

HPLC (Method 4), RT 28.47 min, 97.4 A %; LCMS (ES⁺, m/e=649.0, M⁺+1); ¹HNMR (300 MHz, CD₃OD) δ 7.90 (s, 1H), 6.17 (d, 1H), 4.97 (m, 1H), 4.47(m, 6H), 4.20 (m, 1H), 4.00 (m, 1H), 3.78 (m, 1H), 3.62 (m, 1H), 3.31(d, 3H), 1.44 (t, 3H), 1.35 (m, 3H), 1.23 (m, 6H); ³¹P NMR (121 MHz,CD₃OD) δ 7.52.

HPLC Method 4:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 15%-18%-20%-35%-97%MeCN (0-10-35-45-50 min), 250 nm.

Compound 4b Two Diastereomers Isolated From a Four Isomer Mixture

The nucleoside (B-N3, 0.8 g, 3.1 mmol) was stirred in triethylphosphate(PO(OEt)₃, 15 mL) 1,8-bis(dimethylamino)naphthalene (0.8 g, 3.721 mmol,1.2 eq) was added and the mixture was cooled to −20° C. Phosphorousoxychloride (POCl₃, 0.96 g, 6.26 mmol, 1.3 eq) was added drop-wise andthe mixture was stirred at −20° C. for 10 min. The mixture was allowedto warm to 0° C. and was stirred at this temperature for 30 min. Thismixture was then cooled to −50° C. (dry ice/EtOH) B-2 (0.47 g, 2.79mmol, 0.9 eq) was added followed by triethylamine (TEA, 0.99 mL, 0.713mmol, 2.3 eq). After stirring at −50° C. for 30 minutes, B-4 (1.36 g,0.93 mmol, 3 eq) was added to the mixture followed by4-dimethylaminopyridine (DMAP, 0.94 g, 0.77 mmol, 2.5 eq). The mixturewas allowed to warm to room temperature and was stirred overnight.Petroleum ether was added to the mixture and the precipitated materialwas isolated by vacuum filtration. This material was purified by columnchromatography (silica gel, 200-300 mesh) using MeOH/DCM (0→7%)providing the crude product (570 mg). The crude product was purified bypreparative column chromatography affording compound 4b, diastereomer 1(30 mg, 1.7%) and compound 4b, diastereomer 2 (50 mg, 2.8%).

Compound 4b, Diastereomer 1

HPLC (Method 5), RT 18.81 min, 95.62 A %; LCMS (ES⁺, m/e=580.0, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.73 (d, 1H), 5.95 (s, 1H), 5.76 (d, 1H), 4.98(m, 1H), 4.44 (m, 3H), 4.06 (m, 2H), 3.82 (m, 2H), 3.53 (m, 1H), 1.42(m, 3H), 1.25 (m, 6H), 1.17 (s, 3H); ³¹P NMR (121 MHz, CD₃OD) δ 7.52.

Compound 4b, Diastereomer 2

HPLC (Method 5), RT 21.37 min, 95.9 A %; LCMS (ES⁺, m/e=580.0, M⁺+1); ¹HNMR (300 MHz, CD₃OD) δ 7.77 (d, 1H), 5.98 (s, 1H), 5.84 (d, 1H), 5.02(m, 1H), 4.48 (m, 2H), 4.31 (m, 1H), 4.05 (m, 2H), 3.82 (m, 2H), 3.53(m, 1H), 1.38 (d, 3H), 1.26 (t, 6H), 1.17 (s, 3H); ³¹P NMR (121 MHz,CD₃OD) δ 7.06.

HPLC Method 5:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 7%-15%-30%-97% MeCN(0-25-30-35 min), 250 nm.

Compound 5bx Two Diastereomers Isolated From a Four Isomer Mixture

B-2 was prepared as described herein. B-5 was prepared according to thefollowing scheme:

Preparation of Methyl Isocyanate

Acetic Acid (8.4 g, 139.9 mmol, 1 eq) was dissolved in toluene (75 mL)in a reaction vessel which had been outfitted for distillation.Triethylamine (19.4 mL, 139.9 mmol, 1 eq) was added and the mixture washeated to 70° C. Diphenyl phosphorazidate (50 g, 181.8 mmol, 1.3 eq) wasslowly added drop-wise. After the addition had completed, the mixturewas heated to 110° C. and the product was collected as a solution intoluene. After 40 mL of distillate had been collected, the mixture wascooled to room temperature and the concentration of the methylisocyanate was determined by NMR (2.4 M, 70%).

The amino acid, (C-3, 3.52 g, 16.65 mmol, 0.95 eq) was dissolved inaqueous NaOH (1.75M, 10 mL). Methyl isocyanate (1.00 g. 17.53 mmol, 1eq) was added as a solution in toluene. The mixture was stirred at roomtemperature and was acidified by the addition of aqueous HCl (20%, 35mL). The mixture was heated at reflux temperature for 30 minutes. Whenthe mixture was cooled, the desired product precipitated as a solid. Thesolid C-4 was isolated by filtration, was washed with distilled water,and was dried to provide 3 g (72%) of the desired product.

This material C-4 (4.8 g, 19.3 mmol, 1 eq) was dissolved in toluene (100mL) at 0° C. and AlCl₃ (10.3 g, 77.27 mmol, 4 eq) was added. The mixturewas stirred at room temperature for 15 h. The reaction mixture wasquenched on addition of AcOH and diluted with methanol. The mixture wasconcentrated under reduced pressure and purified by flash columnchromatography (silica gel, petroleum ether: ethyl acetate 10:1 to 1:1)to give B-5 (1.0 g).

The nucleoside (B-N1, 1.5 g, 4.82 mmol) was stirred in triethylphosphate(PO(OEt)₃, 30 mL) and was cooled to 0° C. Phosphorous oxychloride(POCl₃, 0.96 g, 6.26 mmol, 1.3 eq) was added drop-wise and the mixturewas stirred at 0° C. for 3 hours. B-2 (0.724 g, 4.34 mmol, 0.9 eq) wasadded followed by triethylamine (TEA, 1.54 mL, 11.08 mmol, 2.3 eq).After stirring for 1 hour at 0° C., B-5 (1.16 g, 7.23 mmol, 1.5 eq) wasadded to the mixture followed by 1,9-diazabicycloundec-7-ene (DBU, 1.47g, 9.64 mmol, 2 eq). The mixture was allowed to warm to room temperatureand was stirred overnight. Petroleum ether (300 ml) was added to themixture and the precipitated material was isolated by vacuum filtration.This material was purified by flash column chromatography (silica gel)using MeOH/DCM (1→4.5%) providing 5bx (2 g). 5bx was purified twice bypreparative (RP) column chromatography affording compound 5bx,diastereomer 1 (35 mg) and compound 5bx, diastereomer 2 (34 mg).

Compound 5bx, Diastereomer 1

HPLC (Method 6), RT 14.41 min, 95.25 A %; LCMS (ES⁺, m/e=647.1, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.97 (s, 1H), 5.98 (s, 1H), 5.02 (m 1H), 4.52(m, 2H), 4.35 (m, 2H), 4.24 (m, 1H), 4.06 (s, 3H), 3.90 (m, 1H), 3.27 (m1H), 3.14 (m, 1H), 2.93 (s, 3H), 1.36 (m, 3H), 1.24 (d, 6H), 1.00 (s,3H); ³¹P NMR (121 MHz, CD₃OD) δ 34.91.

Compound 5bx, Diastereomer 2

HPLC (Method 6), RT 16.44 min, 95.31 A %; LCMS (ES⁺, m/e=647.1, M⁺+1);¹H NMR (300 MHz, CD₃OD) δ 7.98 (s, 1H), 5.98 (s, 1H), 4.99 (m, 1H), 4.50(m, 4H), 4.22 (m, 1H), 4.06 (s, 3H), 3.95 (m, 1H), 3.42 (m, 1H), 3.22(m, 1H), 2.96 (s, 3H), 1.35 (m, 3H), 1.23 (m, 6H), 0.99 (s, 3H); ³¹P NMR(121 MHz, CD₃OD) δ 34.79.

HPLC Method 6:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 15%-20%-40%-97% MeCN(0-25-30-35 min), 282 nm.

Compound 6 Two Diastereomers Isolated From a Four Isomer Mixture

C2. A solution of trifluoroacetic acid (TFA, 340 mg, 3 mmol, 3 eq) inDCM (10 mL) was added to a solution of the BOC protected amine (C1, 325mg, 1 mmol, 1 eq) in DCM (10 mL) at 0° C. under a nitrogen atmosphere.The mixture was allowed to warm and was stirred at room temperature for1 hour. The mixture was diluted with DCM (20 mL) and was washed with asaturated solution of sodium bicarbonate followed by brine. The organicsolution was dried over sodium sulfate and was concentrated underreduced pressure to provide C2 (270 mg, quant).

The nucleoside (B-N1, 310 mg, 1.00 mmol, 1.0 eq) was stirred intriethylphosphate (PO(OEt)₃, 10 mL). Triethylamine (121 mg, 1.2 mmol,1.2 eq) was added and the solution was cooled to 0° C. under a nitrogenatmosphere. Phosphorous oxychloride (POCl₃, 184 mg, 153 mmol, 1.2 eq)was added drop-wise and the mixture was stirred at 0° C. for 2 hours.Triethylamine (404 mg, 4.0 mmol, 4.0 eq) was added and the mixture wasstirred at 0° C. A solution of C2 (270 mg, 1.2 mmol, 1.2 eq) in DCM (10mL) was added drop-wise, the mixture was allowed to warm to roomtemperature and was stirred overnight. The reaction mixture was pouredinto petroleum ether (100 ml) and the precipitated material was isolatedby vacuum filtration. The precipitate was dissolved in DCM (50 mL), waswashed with brine, and was dried over sodium sulfate. This material waspurified by flash column chromatography (silica gel) using MeOH/DCM(1:30 to 1:20) providing 6 as a mixture of four isomers. The fourisomers were separated by preparative HPLC to provide compound 6,diastereomer 1 (36 mg) and compound 6, diastereomer 2 (43 mg) along witha mixture of the other two diastereomers (42 mg).

Compound 6, Diastereomer 1

HPLC (Method 7), RT 6.69 min, 99.15 A %; LCMS (ES⁺, m/e=581.0, M⁺+1); ¹HNMR (300 MHz, CDCl₃) δ 7.73 (s, 1H), 7.30 (m, 5H), 5.89 (s, 1H), 5.34(br-s, 2H), 5.20 (m, 2H), 4.74 (m, 2H), 4.19-4.32 (m, 4H), 3.97 (s, 3H),3.09 (m, 2H), 2.21 (m, 2H), 1.00 (s, 3H); ³¹P NMR (121 MHz, CDCl₃) δ29.28.

Compound 6, Diastereomer 2

HPLC (Method 7), RT 7.62 min, 98.97 A %; LCMS (ES⁺, m/e=581.0, M⁺+1); ¹HNMR (300 MHz, CDCl₃) δ 7.85 (s, 1H), 7.30 (m, 5H), 6.01 (s, 1H), 5.30(br-s, 2H), 5.13 (m, 2H), 4.88 (m, 1H), 4.48 (m, 2H), 4.42 (m, 3H), 4.02(s, 3H), 3.04 (m, 2H), 2.10 (m, 2H), 0.98 (s, 3H); ³¹P NMR (121 MHz,CDCl₃) δ 31.50.

HPLC Method 7:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 22%-29%-97% MeCN(0-12-17 min), 250 nm.

Compound 7b Two Diastereomers Isolated From a Two Isomer Mixture

L-Pyroglutamic acid (12.9 g, 100 mmol, 1.0 eq) was dissolved in a 1:1mixture of benzyl alcohol and DMF (100 mL). The mixture was cooled to 0°C. and thionyl chloride (15 g, 150 mmol, 1.5 eq) was added drop-wise.The mixture was allowed to warm to room temperature and was stirredovernight. The reaction mixture was concentrated and the crude productwas purified by column chromatography (silica gel) to afford D-1 (10 g,49%).

D-1 (8 g, 36.5 mmol, 1 eq) was dissolved in DCM (150 mL). The mixturewas stirred and was cooled to 0° C. Dimethylaminopyridine (DMAP, 4.9 g,40.1 mmol, 1.1 eq) was added followed by Boc-anhydride ((BOC)₂O, 8.75 g,40.1 mmol, 1.1 eq). The mixture was allowed to warm to room temperatureand was stirred for 2 hours. The mixture was then diluted with DCM (250mL) and was washed successively with aqueous HCl (1N, 50 mL), aqueoussaturated bicarbonate solution (350 mL) and brine. The organic phase wasdried over sodium sulfate, was concentrated under reduced pressure, andwas purified by silica gel chromatography to provide D-2 (8.6 g, 73.8%).LCMS (ES⁺, m/e=656, [2M+NH₄ ⁺]).

A solution of potassium dihydrogen phosphate (K₂H₂PO₄, 2.6 g, 19 mmol,7.3 eq) in a mixture of methanol and water (5:1, 30 mL) was prepared.D-2 (823 mg, 2.6 mmol, 1 eq) was added and the mixture was cooled to 0°C. Sodium borohydride (700 mg, 19 mmol, 7.3 eq) was added and themixture was stirred at 0° C. for 2 hours. Acetic acid (2 mL) was addedand the mixture was stirred for another 20 minutes. The mixture wasfiltered and the filtrate was concentrated under reduced pressure. Theresulting residue was extracted with ether. The ether solution was driedover sodium sulfate, was concentrated under reduced pressure, and theresidue was purified by column chromatography (silica gel) to afford D-3(100 mg, 12%). LCMS (ES⁺, m/e=346, [M+Na]⁺).

D-3 (460 mg, 1.4 mmol, 1 eq) was dissolved in DCM (2.5 mL).Triethylamine (200 mg, 2 mmol, 1.5 eq) was added and the mixture wasstirred at 0° C. under a nitrogen atmosphere. Methane sulfonylchloride(MsCl, 205 mg, 1.8 mmol, 1.2 eq) was added and the mixture was stirredfor 2 hours. The reaction mixture was diluted with DCM and was washedsuccessively with cold aqueous HCl (1N, 5 mL), aqueous saturatedbicarbonate solution (5 ml) and brine (5 mL). The organic phase wasdried over sodium sulfate and was concentrated under reduced pressure.The residue was purified by column chromatography (silica gel) to affordD-4 (350 mg, 49%).

D-4 (5.3 g, 13.2 mmol, 1 eq) was dissolved in THF (40 mL). Potassiumcarbonate (2.1 g, 26.4 mmol, 2.2 eq) was added followed by thioaceticacid (1.2 mL, 26.4 mmol, 2.2 eq). The mixture was allowed to stir atroom temperature overnight. The reaction mixture was diluted with THF(50 mL). Ammonium hydroxide (14.7 M, 30 mL) and water (50 mL) were addedand the mixture was stirred at room temperature for 20 minutes. Themixture was then acidified with HCl (12N) and was extracted with DCM.The organic phase was dried over sodium sulfate and was concentratedunder reduced pressure. The resulting residue was purified by silica gelchromatography to provide D-5 (2.2 g. 49%). LCMS (ES⁺, m/e=677, [M+H]⁺).

D-5 (300 mg, 0.44 mg, 1 eq) was dissolved in DCM (10 mL). Triethylamine(0.18 mL, 1.4 mmol, 3 eq) was added followed by dithiothreitol (DTT,0.28 g, 1.8 mmol, 4 eq). The mixture was stirred at room temperatureunder a nitrogen atmosphere for 2 hours. The reaction mixture wasdiluted with DCM and was washed with 10% citric acid (15 mL) and water(15 mL). The organic phase was dried over sodium sulfate, wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography to afford D-6 (200 mg, 67%).

D-6 (400 mg) was treated with trifluoroacetic acid (3 mL) in DCM (10 mL)at 0° C. to remove the BOC protecting group. This mixture wasconcentrated and the crude material was dissolved in DCM. In a separateflask, phosphorous oxychloride (220 mg. 1.4 mmol, 1 eq) was dissolved inDCM (5 mL). The mixture was cooled to 0° C. and triethylamine (400 mg, 4mmol, 2.8 eq) was added. The solution of the deprotected D-6 in DCMprepared in the first step was added and the mixture was stirred at 0°C. under nitrogen for 2 hours. The mixture was allowed to warm to roomand was stirred for an additional 2 hours. The mixture was concentratedunder vacuum and the residue, D-7, was dissolved in anhydrous THF foruse in the coupling step.

The nucleoside (B-N1, 311 mg, 1 mmol, 1 eq) was dissolved in dry THF (3mL). The mixture was cooled to 0° C., a solution of t-BuMgCl (1M in THF,2 mL, 2 mmol, 2 eq) was added, and the mixture was stirred at 0° C. for2 hours. The solution of D-7 (1.17 mmol) prepared above, was addeddrop-wise. The mixture was allowed to warm to room temperature and wasstirred overnight. The reaction mixture was diluted with DCM (5 mL) andwas washed with water (15 mL) and brine (15 mL). The organic phase wasdried over sodium sulfate, was concentrated under reduced pressure, andwas purified by silica gel column chromatography to provide 7 (30 mg,5%). The crude product was purified by preparative HPLC to providesamples of the two individual diastereomers compound 7b, diastereomer 1(24 mg), and compound 7b, diastereomer 2 (21 mg).

Compound 7b, Diastereomer 1

HPLC (Method 8), RT 8.40 min, 97.32 A %; LCMS (ES⁺, m/e=595.0, M⁺+1); ¹HNMR (300 MHz, CDCl₃) δ 8.05 (s, 1H), 7.65 (m, 5H), 5.88 (s, 1H), 5.35(br-s, 2H), 5.18 (m, 2H), 4.80 (m, 3H), 4.25 (m, 3H), 4.06 (s, 3H) 3.10(m, 1H), 2.91 (m, 1H), 2.25 (m, 2H), 1.38 (m, 2H), 1.00 (s, 3H); ³¹P NMR(121 MHz, CDCl₃) δ 40.12.

Compound 7b, Diastereomer 2

HPLC (Method 8), RT 7.21 min, 99.32 A %; LCMS (ES⁺, m/e=594.9, M⁺+1); ¹HNMR (300 MHz, CDCl₃) δ 7.79 (s, 1H), 7.31 (m, 5H), 5.99 (s, 1H), 5.30(br-s, 2H), 5.10 (m, 2H), 4.86 (m, 1H), 4.57 (m, 1H), 4.40 (m, 2H), 4.25(m, 2H), 4.04 (s, 3H), 3.07 (m, 1H), 2.90 (m, 1H), 1.80-2.23 (m, 4H),0.97 (s, 3H); ³¹P NMR (121 MHz, CDCl₃) δ 41.05.

HPLC Method 8:

C18, 4.6×150 mm, 5 micron, 0.1% Ammonium acetate, 25%-29%-97% MeCN(0-12-17 min), 250 nm.

Compound 8a

Compound 8a was synthesized from 2′-chloro-2′-C-methyl-uridine andD-alanine isopropyl ester.HCl according to the procedure described forcompound 8b in Scheme 1. The mixture of diastereoisomers was separatedby MS-preparative HPLC.

Compound 8a Diastereomer 1

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.25 (d, J=6.22 Hz, 6H),1.41 (d, J=7.16 Hz, 3H), 1.54 (s, 3H), 1.61-1.79 (m, 6H), 2.11-2.18 (m,2H), 3.51-3.59 (m, 1H), 3.87-3.95 (m, 1H), 3.98-4.00 (m, 1H), 4.16-4.19(m, 1H), 4.31 (ddd, J=11.93 Hz, J=5.97 Hz, J=2.61 Hz 1H), 4.51 (ddd,J=12.08 Hz, J=6.12 Hz, J=2.16 Hz, 1H), 5.00 (heptuplet, J=6.26 Hz, 1H),5.76 (d, J=8.13 Hz, 1H), 6.39 (s, 1H), 7.83 (d, J=8.13 Hz, 1H); ³¹P NMR(MeOD, 161.98 MHz) δ (ppm) 34.12 (s, 1P); MS (ESI) m/z=554.2 (MH⁺).

Compound 8a Diastereomer 2

White solid; ¹H NMR (MeOD, 400 MHz) δ (ppm) 1.25 (d, J=6.25 Hz, 3H),1.26 (d, J=6.25 Hz, 3H), 1.42 (d, J=7.18 Hz, 3H), 1.54 (s, 3H),1.61-1.79 (m, 6H), 2.11-2.18 (m, 2H), 3.48-3.54 (m, 1H), 3.92-4.00 (m,1H), 4.02-4.05 (m, 1H), 4.16-4.20 (m, 1H), 4.34 (ddd, J=11.94 Hz, J=6.46Hz, J=2.74 Hz 1H), 4.49 (ddd, J=12.01 Hz, J=5.81 Hz, J=1.89 Hz, 1H),5.00 (heptuplet, J=6.20 Hz, 1H), 5.78 (d, J=8.20 Hz, 1H), 6.41 (s, 1H),7.84 (d, J=8.20 Hz, 1H); ³¹P NMR (MeOD, 161.98 MHz) δ (ppm) 34.18 (s,1P); MS (ESI) m/z=554.2 (MH⁺).

Example 2 HCV Replicon Assay

Huh-7-derived cell line (Zluc) that harbors an HCV genotype 1b repliconand a luciferase reporter gene was grown in Dulbecco's Modified EagleMedium (DMEM) supplemented with 10% fetal bovine serum, 2 mM GlutaMAX,1% MEM nonessential amino acids, 100 IU/mL penicillin, 100 μg/mLstreptomycin, and 0.5 mg/mL Geneticin® (G418). For dose response testingthe cells were seeded in 96-well plates at 7.5×10³ cells per well in avolume of 50 μL, and incubated at 37° C./5% CO₂. Drug solutions weremade up freshly in Huh-7 media as 2× stocks. Ten additional 5-folddilutions were prepared from these stocks in DMEM without G418. At leastthree hours after Zluc cells were seeded, drug treatment was initiatedby adding 50 μL of drug dilutions to the plates in duplicate. Finalconcentrations of drug ranged from 100 μM to 0.0000512 μM. Cells werethen incubated at 37° C./5% CO₂. Alternatively, compounds were tested attwo concentrations (1 μM and 10 μM). In all cases, Huh-7 (which do notharbors the HCV replicon) served as negative control. After 72 hours ofincubation, the inhibition of HCV replication was measured byquantification of photons emitted after mono-oxygenation of5′-fluoroluciferin to oxyfluoroluciferin by firefly luciferase. Forthis, media was removed from the plates via gentle tapping. Fiftymicroliters of ONE-glo luciferase assay reagent was added to each well.The plates were shaken gently for 3 min at room temperature andluminescence was measured on a Victor³ V 1420 multilabel counter (PerkinElmer) with a 1 second read time using a 700 nm cut-off filter. The EC₅₀values were calculated from dose response curves from the resultingbest-fit equations determined by Microsoft Excel and XLfit 4.1 software.When screening at two fixed concentrations, the results were expressedas % inhibition at 1 μM and 10 μM.

For cytotoxicity evaluation, Zluc cells were treated with test compoundas described herein, and cell viability was monitored using theCellTiter-Blue Cell Viability Assay (Promega) by adding 20 μL of theassay solution to each well. The plates were then incubated at 37° C./5%CO₂ for at least 3 hours. Fluorescence was detected in plates usingexcitation and emission wavelengths of 560 and 590 nm, respectively, ina Victor³ V 1420 multilabel counter (Perkin Elmer) and CC₅₀ values weredetermined using Microsoft Excel and XLfit 4.1 software.

Compounds presented in Table 1 below were assayed according to thereplicon assay described herein.

TABLE 1 HCV Replicon Activity Compound HCV Replicon Compound HCVReplicon Reference EC₅₀ CC₅₀ Reference EC₅₀ CC₅₀ Compound 1b ++ ++Compound 1b + ++ Diastereomer 1 Diastereomer 2 Compound 23b ++ ++Compound 23b ++ ++ Diastereomer 1 Diastereomer 2 Compound 3b ++ ++Compound 3b ++ ++ Diastereomer 1 Diastereomer 2 Compound 4b + ++Compound 4b + ++ Diastereomer 1 Diastereomer 2 Compound 8a + ++ Compound8a + ++ Diastereomer 1 Diastereomer 2 Compound 8b ++ ++ Compound 8b +++++ Diastereomer 1 Diastereomer 2 Compound 5bx ++ + Compound 5bx ++ +Diastereomer 1 Diastereomer 2 Compound 6 ++ ++ Compound 6 ++ ++Diastereomer 1 Diastereomer 2 Compound 7b + ++ Compound 7b ++ ++Diastereomer 1 Diastereomer 2 Compound 9b ++ ++ Compound 9b ++ ++Diastereomer 1 Diastereomer 2 Compound 12b + ++ Compound 12b ++ ++Diastereomer 1 Diastereomer 2 Compound 11b ++++ ++ Compound 11b ++++ ++Diastereomer 1 Diastereomer 2 Compound 14b + ++ Compound 14b ++ ++Diastereomer 1 Diastereomer 2 Compound 16b + ++ Compound 16b + ++Diastereomer 1 Diastereomer 2 Compound 15b + ++ Compound 2by ++ ++Compound 10b ++++ ++ Compound 13b ++ ++ Compound 17b + ++ EC₅₀ isprovided as follows: ++++ ≦ 250 nM, 250 nM < +++ ≦ 1 μM, 1 μM < ++ ≦ 10μM, 10 μM < + CC₅₀ is provided as follows: + ≦ 50 μM, 50 μM < ++

All publications, patents, and patent applications cited in thisspecification are herein incorporated by reference as if each individualpublication, patent, or patent application were specifically andindividually indicated to be incorporated by reference. While theclaimed subject matter has been described in terms of variousembodiments, the skilled artisan will appreciate that variousmodifications, substitutions, omissions, and changes may be made withoutdeparting from the spirit thereof. Accordingly, it is intended that thescope of the claimed subject matter is limited solely by the scope ofthe following claims, including equivalents thereof.

What is claimed is:
 1. A compound according to Formula I:

or a pharmaceutically acceptable salt, solvate, stereoisomeric form,tautomeric form, or polymorphic form thereof, wherein: Base is anucleobase; R^(A) is hydroxyl or fluoro; R^(B) is methyl or halo; PD is

X is alkyl, alkoxycarbonylalkyl,(alkoxycarbonyl)(alkoxycarbonylamino)alkyl, cycloalkyl,heterocycloalkyl, hydantoinylalkyl, or aryl; Y is —NR¹R² or an N-linkedamino acid, or ester thereof; Z is alkyl, alkoxycarbonyl oralkoxyalkylcarbonyl; A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or —CHR¹⁰CHR¹¹CHR¹²—;R¹ is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl, orheteroarylalkyl; R² is hydrogen or alkyl; R is hydrogen or alkyl; andeach of R¹⁰, R¹¹, and R¹² is independently hydrogen or alkyl; or R¹⁰ andR¹¹, together with the carbon atoms to which they are attached, combineto form a five-, six-, or seven-membered ring.
 2. The compound of claim1 according to Formula III:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 3. The compound ofclaim 1 according to Formula II:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 4. The compound ofclaim 1 according to any of Formulas IV-VI:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 5. The compound ofclaim 1 according to Formula VII or VIII:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 6. The compound ofclaim 1, wherein Base is:

or tautomeric form thereof, wherein: R⁴ is hydrogen, hydroxyl,unsubstituted alkoxyl, —NR^(1′)R^(2′) or alkyl substituted with—NR^(1′)R^(2′); where each R^(1′) and R^(2′) are independently selectedfrom hydrogen, unsubstituted alkyl and unsubstituted cycloalkyl; R⁵ ishydrogen, hydroxyl, —NR^(1′) R^(2′) (where R^(1′) and R^(2′) areindependently selected from hydrogen, unsubstituted alkyl andunsubstituted cycloalkyl), or unsubstituted alkoxyl; R⁶ is hydrogen,halogen, or unsubstituted alkyl; and R⁷ is hydrogen or —NH₂.
 7. Thecompound of claim 6 according to any of Formulas (IX)-(XII):

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 8. The compound ofclaim 1 according to any of Formulas XIII-XVII:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 9. A compoundaccording to any of Formulas 1-23:

or a pharmaceutically acceptable salt, solvate, stereoisomericform,-tautomeric form, or polymorphic form thereof.
 10. A pharmaceuticalcomposition comprising the compound of claim 1 and a pharmaceuticallyacceptable excipient, carrier, or diluent.
 11. The pharmaceuticalcomposition of claim 10, wherein the composition is an oral formulation.12. A method for the treatment of a host infected with a hepatitis Cvirus, comprising the administration of an effective treatment amount ofa compound of claim
 1. 13. The method of claim 12, wherein the host is ahuman.
 14. The method of claim 12, wherein the administration directs asubstantial amount of the compound, or pharmaceutically acceptable saltor stereoisomer thereof, to a liver of the host.
 15. The method of claim12, wherein the compound is administered in combination or alternationwith a second anti-viral agent, wherein the second anti-viral agent isan interferon, a nucleotide analogue, a polymerase inhibitor, an NS3protease inhibitor, an NS5A inhibitor, an entry inhibitor, anon-nucleoside polymerase inhibitor, a cyclosporine immune inhibitor, anNS4A antagonist, an NS4B-RNA binding inhibitor, a locked nucleic acidmRNA inhibitor, a cyclophilin inhibitor, or combination thereof.
 16. Themethod of claim 15, wherein the second anti-viral agent is IDX-719,TMC435, PSI-7977, telaprevir, boceprevir, interferon alfacon-1,interferon alfa-2b, pegylated interferon alpha 2a, pegylated interferonalpha 2b, ribavirin, and combinations thereof.
 17. The method of claim15, wherein the second anti-viral agent is IDX-719, TMC435, PSI-7977,telaprevir, boceprevir, interferon alfacon-1, interferon alfa-2b,pegylated interferon alpha 2a, pegylated interferon alpha 2b, andcombinations thereof, and further wherein the administration is not incombination or alternation with ribavirin.
 18. The compound of claim 1,wherein: Base is

or a tautomeric form thereof; R⁴ is hydrogen, hydroxyl, unsubstitutedalkoxyl, —NR^(1′)R^(2′) or alkyl substituted with —NR^(1′)R^(2′); whereeach R^(1′) and R^(2′) are independently selected from hydrogen,unsubstituted alkyl and unsubstituted cycloalkyl; R⁵ is hydrogen;hydroxyl; —NR^(1′)R^(2′) where R^(1′) and R^(2′) are independentlyselected from hydrogen, unsubstituted alkyl and unsubstitutedcycloalkyl; or unsubstituted alkoxyl; R⁶ is hydrogen, halogen, orunsubstituted alkyl; and R⁷ is hydrogen or —NH₂; R^(A) is hydroxyl orfluoro; R^(B) is methyl or halo; PD is

X is unsubstituted alkyl; substituted alkyl; alkoxycarbonylalkyl;(alkoxycarbonyl)(alkoxycarbonylamino)alkyl; cycloalkyl;heterocycloalkyl;

where R^(XX) and R^(YY) each independently hydrogen or unsubstitutedlower alkyl; or aryl; Y is —NR¹R², —NR^(X)-G(S_(C))—C(O)-Q¹, or—O—C(O)-G(S_(C))—NH-Q²; R¹ is hydrogen, unsubstituted alkyl, substitutedalkyl, cycloalkyl, aryl, heteroaryl, -alkyl-(unsubstituted aryl),-alkyl-(substituted aryl), or heteroarylalkyl; R² is hydrogen orunsubstituted alkyl; Q¹ is —SR^(Y), —NR^(Y)R^(Y) or unsubstitutedalkoxyl; R^(Y) is hydrogen or unsubstituted alkyl; S_(C) is a side chainof a naturally occurring or non-naturally occurring amino acid; or S_(C)is hydrogen, unsubstituted alkyl, substituted alkyl,-alkyl-(unsubstituted aryl), -alkyl-(substituted aryl),heterocycloalkyl, carboxylalkyl, heteroarylalkyl, alkyl substituted with—NR^(1′)R^(2′) (where each R^(1′) and R^(2′) are independently selectedfrom hydrogen, unsubstituted alkyl and unsubstituted cycloalkyl),hydroxylalkyl, -alkyl-NH—C(NH)—NH₂, -alkyl-C(O)NH₂, sulfanylalkyl,alkylsulfanylalkyl, or hydroxylarylalkyl; G is C₁-C₂ alkyl; R^(X) ishydrogen or R^(X) and S_(C), together with the atoms to which they areattached, combine to form a five-membered heterocyclic ring; Q² ishydrogen or unsubstituted alkoxyl; or Q² and S_(C), together with theatoms to which they are attached, combine to form a five-memberedheterocyclic ring; Z is unsubstituted alkyl, substituted alkyl,alkoxycarbonyl or alkoxyalkylcarbonyl; A is —CHR¹⁰—, —CHR¹⁰CHR¹¹—, or—CHR¹⁰CHR¹¹CHR¹²—; R is hydrogen or unsubstituted alkyl; each of R¹⁰,R¹¹, and R¹² is independently hydrogen or unsubstituted alkyl; or R¹⁰and R¹¹, together with the carbon atoms to which they are attached,combine to form a five-, six-, or seven-membered ring; each “alkyl,”unless specified otherwise, is unsubstituted; each alkoxy is —OR′ whereR′ is unsubstituted alkyl or unsubstituted cycloalkyl; each “substitutedalkyl” is independently alkyl substituted with moiety(ies) selected fromhalogen; hydroxyl; sulfanyl; —NR^(1′)R^(2′) where each R^(1′) and R^(2′)are independently selected from hydrogen, unsubstituted alkyl andunsubstituted cycloalkyl; —NH (unsubstituted aryl), —N(unsubstitutedaryl)₂; —OR′ where R′ is unsubstituted alkyl or unsubstitutedcycloalkyl; —O-(unsubstituted aryl); nitro; cyano; sulfonic acid;sulfate; phosphonic acid; and phosphate; each cycloalkyl is a saturated,monocyclic hydrocarbon or a bridged or fused bicyclic hydrocarboncomprising 3 to 15 carbon atoms; the cycloalkyl is unsubstituted orsubstituted, unless otherwise specified, and when substituted issubstituted with moiety(ies) selected from halogen; hydroxyl; sulfanyl;—NR^(1′)R^(2′) where each R^(1′) and R^(2′) are independently selectedfrom hydrogen, unsubstituted alkyl and unsubstituted cycloalkyl;—NH(unsubstituted aryl), —N(unsubstituted aryl)₂; —OR′ where R′ isunsubstituted alkyl or unsubstituted cycloalkyl; —O-(unsubstitutedaryl); nitro; cyano; sulfonic acid; sulfate; phosphonic acid; andphosphate; the heterocyclo in heterocycloalkyl is a monovalent,monocyclic, non-aromatic ring system or a fused or bridged, bicyclic,tricyclic, or tetracyclic ring system that is partially or fullysaturated or aromatic provided that it contains at least onenon-aromatic ring; wherein the heterocyclo comprises 3 to 20 ring atoms;wherein one or more of the non-aromatic ring atoms are heteroatomsindependently selected from O, S, or N and the remaining ring atoms arecarbon atoms and where the nitrogen or sulfur atoms may be optionallyoxidized, and the nitrogen atoms may be optionally quaternized; andwherein the heterocyclo may be attached to the alkyl at any heteroatomor carbon atom which results in the creation of a stable compound; andwherein the heterocyclo ring is unsubstituted or substituted and whensubstituted is substituted with moiety(ies) selected from halogen;hydroxyl; alkoxycarbonyl; alkoxycarbonylalkyl; sulfanyl; —NR^(1′)R^(2′)where each R^(1′) and R^(2′) are independently selected from hydrogen,unsubstituted alkyl and unsubstituted cycloalkyl; —NH(unsubstitutedaryl), —N(unsubstituted aryl)₂; —OR′ where R′ is unsubstituted alkyl orunsubstituted cycloalkyl; —O-(unsubstituted aryl); nitro; cyano;sulfonic acid; sulfate; phosphonic acid; and phosphate; each aryl is aC₆-C₁₂ aromatic group and is unsubstituted or substituted, unlessotherwise specified, and when substituted is substituted withmoiety(ies) selected from halogen; unsubstituted alkyl; substitutedalkyl; haloalkyl; hydroxyl; —NR^(1′)R^(2′) where each R^(1′) and R^(2′)are independently selected from hydrogen, unsubstituted alkyl andunsubstituted cycloalkyl; —NH(unsubstituted aryl), —N(unsubstitutedaryl)₂; —OR′ where R′ is unsubstituted alkyl or unsubstitutedcycloalkyl; —O-(unsubstituted aryl); nitro; cyano; sulfonic acid;sulfate; phosphonic acid; and phosphate; and each heteroaryl is amonovalent monocyclic aromatic group or multicyclic aromatic group thatcontains at least one aromatic ring; wherein the ring comprises 5 to 20ring atoms; wherein at least one aromatic ring contains one to fourheteroatoms independently selected from O, S, and N, provided that thering contains at least one carbon atom; and where the heteroaryl isbonded to the rest of the molecule through the aromatic ring.
 19. Apharmaceutical composition comprising the compound of claim 18 and apharmaceutically acceptable excipient, carrier, or diluent.
 20. A methodfor the treatment of a host infected with a hepatitis C virus,comprising the administration of an effective treatment amount of acompound of claim 18.